Chromophore assignment in C-phycocyanin from ... - Core

ZEITSCHRIFT FÜR N A T U R F O R S C H U N G

Information for Contributors Contributions to Z . Naturforsch., Section C (biological sciences), may be (i) original papers, (ii) research notes not exceeding two pages in print and (iii) reports on current research of special interest. Two copies of each contribution should be sent to Dr. Helga Hausen, P.O. Box 2645, D-7400 Tübingen. (Section B (chemical sciences): Dr. Heide Voelter, P.O. Box 2645, D-7400 Tübingen; Section A (physical sciences): Mrs. Tamara Littmann, Lenzhalde 21, D-7082 Oberkochen). Contributors are encouraged to suggest possible reviewers of their papers. The manuscript should be type written using double spacing throughout. The title should be concise but informative. The names of the authors, with first names and initials, and of the institution where the work was carried out follow the title. A running title with not more than 60 characters should be indicated if the title is longer than this. In a footnote on the first page, please give an address for reprint requests. The title, a selfconsistant abstract, suitable for direct use by the abstracting journals (no references to the main text), and five keywords, all in English, must precede the main text of each contribution. The main text should meet the highest standards as to novelty of the material, organization and conciseness. A qualified colleague and, if the text is in a foreign language, a person who thoroughly knows it, should have been given the opportunity to check the paper before its submission for publication. Papers, reporting mainly spectroscopic, X-ray or other data of no general relevancy should not be submitted. References and footnotes should be numbered (e.g. [1]) and listed at the end of the paper (e.g. [1] A . Meyer, Z . Naturforsch. 30c, 633 (1976)). Tables with the appropriate captions and a list of the figure legends should follow at the end of the paper. Allowance should be made for the reduction in printing of the drawings (line thickness, lettering!). Original drawings larger than 21 x 30cm should be replaced by copies of reduced size, when the manuscript is submitted and only be sent in when the manuscript is accepted. On all illustrations, the figure number and the author's name must be written in pencil. The authors will receive page proofs. Changes in the text after acceptance of the paper and drawings not fit for direct reproduction cause delay and create extra costs which may be charged to the author. 50 reprints are free of charge.

Section C A Journal of Biosciences ISSN 0341-0382

Managing Editor: H . Hausen, Tübingen Council: E. Bünning, Tübingen A . Butenandt, München M . Eigen, Göttingen

Editors: A . Hager, Tübingen K. Hahlbrock. Köln W. Hasselbach, Heidelberg P. Karlson, Marburg F. Kaudewitz, München J. Klein, Tübingen J. St. Schell, Köln E. Wecker, Würzburg

Associate Editors: N . Amrhein, Bochum B. A . Askonas, London W. Barz, Münster P. Böger, Konstanz G. Bornkamm, Freiburg D. Bückmann, Ulm K. G. Götz, Tübingen G. Gottschalk, Göttingen P. Gruss, Heidelberg G. Isenberg, Köln R. Jaenicke, Regensburg V. ter Meulen, Würzburg G. F. Meyer, Tübingen M . Rajewsky, Essen H . Schimassek, Heidelberg D. Schulte-Frohlinde, Mühlheim/R. G. Schulz, Freiburg F. F. Seelig, Tübingen J. Seelig, Basel H . Simon, München W. Steglich, Bonn H . Stieve, Aachen J. Suko, Wien A . Trebst, Bochum G. Weissenböck, Köln G. Wiek, Innsbruck V . Zimmermann, Würzburg

Information for Subscribers The subscription prices per year are for normal subscribers: Section A DM510.-

Section B DM610.-

Section C D M 425.-

for authors and their institutions if they order directly from the publishers: Section A D M 408. -

Section B D M 488.-

Section C D M 340. -

plus postage and handling Section A DM35.-

Section B DM35.-

Section C DM25.-

Single copies and back-numbers are available. Subscriptions will remain standing for the following year unless cancellations are made by postage October 1st. Informations for Advertisers The price for a small advertisement (breadth 43 mm, height 57 mm) in all three sections of the journal is D M 6 0 . - . For larger advertisments please order the price list from the publishers.

V E R L A G D E R ZEITSCHRIFT F Ü R NATURFORSCHUNG, T Ü B I N G E N P.O. Box 2645, D-7400 Tübingen (Postscheck-Konto Stuttgart 8039-700).

I

Bayerische

]

Z E I T S C H R I F T FÜR NATURFORSCHUNG SECTION C A EUROPEAN JOURNAL O F

BIOSCIENCES Council

Editorial Board

E . BüNNiNG,Tübingen A . BUTENANDT, München M. EIGEN, Göttingen

A. H A G E R , T ü b i n g e n K. HAHLBROCK, Köln W . H A S S E L B A C H , Heidelberg P. K A R L S O N , M a r b u r g F. K A U D E W I T Z , M ü n c h e n J. K L E I N , T ü b i n g e n J. ST. S C H E L L , K ö l n E. W E C K E R , W ü r z b u r g

Advisory Editorial Board N . A M R H E I N , Bochum B. A . ASKONAS, London W. BARZ, Münster P . BÖGER, Konstanz G . B O R N K A M M , Freiburg D.

BÜCKMANN, Ulm

K. G. GÖTZ, Tübingen G . GOTTSCHALK, G ö t t i n g e n P . GRUSS, Heidelberg

G. R. V. G. M. H.

ISENBERG, K ö l n J A E N I C K E , Regensburg TER M E U L E N , W ü r z b u r g F. MEYER, Tübingen R A J E W S K Y , Essen SCHIMASSEK, Heidelberg

D.

SCHULTE-FROHLINDE,

EDITED WITH

Mühlheim/R.

G . S C H U L Z , Freiburg F. F . SEELIG, T ü b i n g e n

T H EINSTITUTES

J. S E E L I G , Basel H . SIMON, München W . STEGLICH, Bonn H . STIEVE, A a c h e n J. S U K O , W i e n A . TREBST, B o c h u m G. WEISSENBÖCK, Köln G . W I C K , Innsbruck V. ZIMMERMANN, Würzburg

I N C O L L A B O R A T I O N O F T H E MAX-PLANCK-GESELLSCHAFT

1987

V o l u m e 42 c

VERLAG DER ZEITSCHRIFT FÜR NATURFORSCHUNG TÜBINGEN

Anschrift des Verlages: Postfach 26 45, D-7400 Tübingen Satz und Druck: Allgäuer Zeitungsverlag GmbH, Kempten

Nachdruck — auch auszugsweise — nur mit schriftlicher Genehmigung des Verlages

Section a Physics, Physical Chemistry, Cosmic Physics Section b Inorganic and Organic Chemistry

1987

Zeitschrift für Naturforschung

Volume 42c

Contents Contents of Number Original

Co mm

Transformations of /rarcs-2-Hexenal by Botrytis cinerea PERS. as Detoxification Mechanisms ( I n German)

1/2

imications

I . URBASCH

X-Ray Structure Analysis and Spectroscopic D a t a of the A n t i b i o t i c 8-(Dichloroacetyl)-5-hydroxy-2,7dimethyl-l,4-naphthoquinone from the Fungus Mollisia sp. G.

WEBER, T.

HÜBNER, A.

G I E R E N , J.

SONNEN-

BICHLER, T . K O W A L S K I , and O. H O L D E N R I E D E R

Alkaloids i n Stem Roots o f Nicotiana Spartium junceum Transformed by rhizogenes M . W I N K and L . W I T T E

Flavonoids f r o m the Leaf Gutierrezia sarothrae

H . R E M B O L D , H . FORSTER, and J. S O N N E N B I C H L E R

4

RADDI

G . SKOPP, H . - J . O P F E R K U C H , and G . S C H W E N K E R 7

Methylation o f Guanine in vivo by the Organophosphorus Insecticide Methamidophos S. M . A . D . Z A Y E D and F. M . M A H D I

17

Cytogenetic Effects o f the Insecticide M e t h a m i d o phos in Mouse B o n e M a r r o w and C u l t u r e d Mouse Spleen Cells S. M . A M E R and M . A . S A Y E D

21

Molluscicidal Properties of Quinones ( I n German) H . S C H I L D K N E C H T and J. L U B O S C H

tabacum and Agrobacterium 69

1

Structure o f A z a d i r a c h t i n B

n-Alkylphenols f r o m Schinus terebinthifolius (Anacardiaceae) ( I n German)

64

31

Resin o f

D . H R A D E T Z K Y , ' E . W O L L E N W E B E R , and J . N .

W . FÖRSTER and

H . BECKER

73

The 1 4 ß - H y d r o x y l a t i o n i n the Biosynthesis o f Cardenolides in Digitalis purpurea. The Role of 3 ß Hydroxy-5ß-pregn-8(14)-en-20-one M . E . D E L U C A , A . M . S E L D E S , and E . G . G R O S

G . F. K O L A R and M . S C H E N D Z I E L O R Z

41

Interaction in vitro o f N o n - E p i t h e l i a l Intermediate Filament Proteins w i t h Histones T R A U B , G.

BARTH

P E R I D E S , S. K Ü H N , and

77

Biotransformation of H u m u l e n e by Fungi and Enantioselectivity of the Strains Used W . - R . A B R A H A M and B . S T U M P F

79

Enzymatic Synthesis o f 1,6-Digalloylglucose from ß Glucogallin by ß - G l u c o g a l l i n : ß - G l u c o g a l l i n 6-0Galloyltransferase from O a k Leaves S.

W . SCHMIDT, K . D E N Z E L , G.

SCHILLING,

and 87

PAPS-Reductase from Escherichia coli: Characterization of the Enzyme as Probe for Thioredoxins J. D . S C H W E N N and U . SCHRIEK

93

33

Reactions o f Substituted A r e n e d i a z o n i u m Chlorides w i t h Methylamine-Formaldehyde Premix Revisited: Reactivity and Transformations of M e t h y l o l amine Intermediates and T h e i r Biological Significance

P.

ROIT-

MAN

G . G . GROSS

Effects of Plant Bioregulators on the Production of I r i d o i d D e r i v e d Terpenoids i n Valeriana wallichii and Fedia cornucopiae Cell Suspension Cultures

Snakeweed,

A.

SCHER47

Protein Transport in Chloroplasts: A T P is Prerequisit C. S C H I N D L E R , R. H R A C K Y , and J. S O L L

103

L i p i d Influence on the Structure of the Light H a r vesting B 8 0 0 - 8 5 0 Proteins J. P E S C H K E and H . M O H W A L D

109

Daffodil Chromoplast D N A : Comparison w i t h C h l o roplast D N A , Physical M a p , and Gene Localization P. H A N S M A N N

118

IV

Contents

U p t a k e of Protoporphyrin and V i o l e t L i g h t Photodestruction of Propionibacterium acnes T. B . M E L 0

123

Species and Sex Specificity in the O d o u r Composition o f T w o Panurgine Bees ( H y m e n o p t e r a , A n drenidae) W.

E v o l u t i o n of E. coli t R N A f r o m Other t R N A s

I l e

: Evidence of D e r i v a t i o n

FRANCKE, W.

SCHRÖDER, A . K . B O R G - K A R L S -

S O N , G . BERGSTRÖM, and J. T E N G Ö

169

M . P. STAVES, D . P. B L O C H , and J. C. L A C E Y , Jr.

129 Intermolecular H - A b s t r a c t i o n of T h i y l Radicals from Thiols and the Intramolecular Complexing of the T h i y l Radical w i t h the T h i o l G r o u p i n 1,4-Dithiot h r e i t o l . A Pulse Radiolysis Study M . S H . A K H L A Q and C. V O N SONNTAG

134

T h e Contents of the D u f o u r G l a n d o f the A n t Harpagoxenus sublaevis N y l . (Hymenoptera: F o r m i cidae) D . G. OLLETT, E. D . M O R G A N , A . B . ATTYGALLE, and J. P. J. B I L L E N

141

Contents of Number 3 Original

Commutations

Analysis o f Single Channel Currents w i t h a M i c r o processor Based Device W.

SCHREIBMAYER,

L U E G E R , and H . A .

E.

HOFER,

P.

WOLF,

TRITTHART

Natural Waxes Investigated by Soft Ionization Mass Spectrometry H.-R.

S C H U L T E N , K . E . M U R R A Y , and N . S I M M L E I T

A G C / M S Study of the Propolis Phenolic Constituents V.

BANKOVA,

A.

DYULGEROV,

S.

POPOV,

N . MAREKOV

and 147

178

Epicuticular Leaf Wax o f Euphorbia Euphorbiaceae P.-G.

Investigation o f the Screening Pigment System in the C o m p o u n d Eye of the M o t h Agrotis segetum (fam. Noctuidae) by Visible Reflectometry T . N O R D T U G and T . B . M E L 0

152

dendroides

G Ü L Z , H . H E M M E R S , J. B O D D E N , and

MARNER

L.. F.-J. 191

Cell-Free Synthesis of the A l k a l o i d s A m m o d e n d r i n e and Smipine M . W I N K and L . W I T T E

197

Comparison of the Solid State C P M A S and Solution C a r b o n - 1 3 - N M R Spectra of H u m i c Acids Extracted from Composted M u n i c i p a l Refuse

Notes Epicuticular Leaf Wax of Cistus albanicus, T . V O G T and P.-G.

Cistaceae

GÜLZ

157

Isopulegol from L i q u i d Cultures o f the Fungus tocystis coerulescens (Ascomycotina) W.-G.

K O C H and V . S I N N W E L L

Cera159

I n h i b i t i o n of cAMP-Phosphodiesterase date

by M o l y b -

B . O F E N L O C H - H Ä H N L E and K . E I S E L E

162

Polyenic Hydrocarbons as Sex Attractants for Geometrids and A m a t i d s (Lepidoptera) Found by Field Screening in Hungary G.

A. 173

Szöcs,

M.

TÖTH,

H.

J.

BESTMANN,

O.

V O S T R O W S K Y , R. R. H E A T H , and J. H . T U M L I N S O N

165

R.

F R Ü N D , F.

J.

GONZALEZ-VILA,

H.-D.

M A N N , and F. M A R T I N

LÜDE205

Glutamic A c i d - l - s e m i a l d e h y d e , a H y p o t h e t i c a l I n termediate in the Biosynthesis o f 5 - A m i n o levulinic A c i d A . K A H and D . D Ö R N E M A N N

Interaction of Sodium, L i t h i u m , Caesium, Potassium Ions w i t h A s c o r b y l Radicals P. W I E C Z O R E K , T . O G O N S K I , and Z . M A C H O Y

209

and 215

The Effect o f G u a n i d i n i u m Chloride on the SelfAssociation o f Bovine L i v e r Glutamate D e h y d r o genase: a G e l Filtration Study A . M A Z Z I N I and R. F A V I L L A

217

V

Contents Molecular Mechanics Investigation on Side-Chain Conformations of a 17a-Ethyl-17ß-hydroxy Steroid w i t h Regard to Receptor B i n d i n g M . BOHL

221

Characterization and A m i n o A c i d Composition o f a Hypertrehalosaemic Neuropeptide from the Corpora cardiaca o f the Cockroach, Nauphoeta cinerea G. G Ä D E

Chains: I n h i b i t o r y Properties vs Purified E. U r i d i n e Phosphorylase A . K . DRABIKOWSKA, L . LISSOWSKA, M .

coli

DRAMIN-

S K I , A . Z G I T - W R O B L E W S K A , and D . S H U G A R

288

A Simple Analysis of Purine and Pyrimidine N u cleotides in Plant Cells by High-Performance L i quid Chromatography H . A S H I H A R A , K . M I T S U I , and T . U K A J I

297

225

The Rate of A T P Hydrolysis Catalyzed by Reconstituted CFoFpLiposomes G . S C H M I D T and P. G R Ä B E R

Magnetotactic Bacteria from Freshwater M . O B E R H A C K , R. S Ü S S M U T H , and H . F R A N K

300

231

Phosphate A c c u m u l a t i o n by Muscle in vitro and the Influence of V i t a m i n D Metabolites 3

T . B E L L I D O and

R. B O L A N D

237

A Correlation between Detergent Tolerance Cell W a l l Structure i n Green Algae

and

Contents of Number 4 Original

Communications

S. B I E D U N G M A I E R , G . W A N N E R , and A . S C H M I D T

245 Further Characterization o f Chickpea Isoflavone 70-Glucoside-6"-0-malonate: malonylesterase: Evidence for a Highly Specific, M e m b r a n e - B o u n d Enzyme in Roots of Cicer arietinum L . W . H I N D E R E R , J. K Ö S T E R , and W . B A R Z

Chromophore Assignment in C-Phycocyanin Mastigocladus laminosus

251

from

S. S I E B Z E H N R Ü B L , R. F I S C H E R , and H . SCHEER

Stereochemistry and Mechanism of Reactions Catalyzed by Tyrosine Phenol-Lyase from Escherichia intermedia M.

M.

PALCIC,

K U M A G A I , S.

S.-J.

SHEN,

SAWADA, H .

E.

SCHLEICHER,

YAMADA,

FLOSS

and

H.

H. G. 307

Distant Precursors o f Benzylisoquinoline A l k a l o i d s and Their Enzymatic F o r m a t i o n M . R U E F F E R and M . H . Z E N K

319

258

Characterization of 2ß( R)-17-0-Acetylajmalan: Acetylesterase — a Specific Enzyme I n v o l v e d i n the Biosynthesis o f the Rauwolfia A l k a l o i d A j maline J

The Effect of Phosphinothricin (Glufosinate) on Photosynthesis. I . I n h i b i t i o n o f Photosynthesis and A c c u m u l a t i o n of A m m o n i a A . W I L D , H . S A U E R , and W . R Ü H L E

263

The Effect o f Phosphinothricin (Glufosinate) on Photosynthesis. I I . The Causes o f I n h i b i t i o n o f Photosynthesis H . S A U E R , A . W I L D , and W . R Ü H L E

270

Sethoxydim-Uptake by Leaf Slices of Sethoxydim Resistant and Sensitive Grasses I . STRUVE, B . G O L L E , and U . L U T T G E

Light-Induced C a

2 +

Influx into Spinach Protoplasts

K . H E I M A N N , G . K R E I M E R , M . M E L K O N I A N , and LATZKO

279

E. 283

L . P O L Z , H . S C H Ü B E L , and J. STÖCKIGT

333

Induction and Characterization o f a N A D P H - D e pendent Flavone Synthase from Cell Cultures o f Soybean G . K O C H S and H . G R I S E B A C H

343

Proposal for the Mechanism o f A c t i o n o f Urocanase. Inference from the I n h i b i t i o n by 2 - M e t h y l u r o c a nate E . G E R L I N G E R and J. R £ T E Y

349

Re-Investigation of the Protein Structure o f Coenzyme B - D e p e n d e n t D i o l Dehydrase 12

Acyclonucleoside Analogues Consisting of 5- and 5,6-Substituted Uracils and Different Acyclic

K . T A N I Z A W A , N . N A K A J I M A , T. TORAYA, H . T A N A K A , and K . S O D A

353

Contents

VI Methanogenesis barkeri:

from

Acetate

Catalysis

by

o f Acetate

Methanosarcina Formation

from

M e t h y l I o d i d e , C 0 , and H by the Enzyme Sys2

2

tem I n v o l v e d

Functional G r o u p Recognition of Pheromone M o l e cules by Sensory Cells of Antheraea polyphemus and Antheraea pernyi (Lepidoptera: Saturniidae) H.

K . L A U F E R , B . E I K M A N N S , U . F R I M M E R , and

R. K .

THAUER

J.

BESTMANN, W . C A I - H O N G , B.

DÖHLA, L I -

K E D O N G , and K . E . K A I S S L I N G

435

360

Divergent E v o l u t i o n of 5S r R N A Genes i n Methanococcus G . W I C H , L . S I B O L D , and A . B Ö C K

373

Characterization o f Some Claviceps Strains D e r i v e d from Regenerated Protoplasts B . S C H U M A N N , W . M A I E R , and D . G R Ö G E R

381

Phenylalanine and Tyrosine Biosynthesis in Sporeforming Members of the O r d e r Actinomycetales H . - K . H U N D , B . K E L L E R , and F. L I N G E N S

B u i l d i n g Blocks for Oligonucleotide Syntheses w i t h U n i f o r m l y Fragmentable ß - H a l o g e n a t e d Protecting Groups ( I n German) P. L E M M E N , R. K A R L , I . U G I , N . B A L G O B I N , and CHATTOPADHYAYA

J. 442

Experiments on the Optical Resolution o f Conduramine Analogs by Enzymatic Transesterification in Organic Solvents ( I n G e r m a n ) G . K R E S Z E and M . S A B U N I

446

387

Steric Course of the Rhodium-Catalyzed Decarbonylation of Chiral 4 - M e t h y l - [ l - H , 2 - H ] p e n t a n a l 3

E. coli M a l t o d e x t r i n Phosphorylase: Primary Structure and Deletion M a p p i n g o f the C-Terminal Site

2

1

H . O T S U K A and H . G . F L O S S

449

D . P A L M , R. G O E R L , G . W E I D I N G E R , R. Z E I E R , B . FISCHER, and

R. S C H I N Z E L

394

Fermentation of D-Xylose to E t h a n o l by

Bacillus

E . B A Y E R , H . H E L L S T E R N , and H . E C K S T E I N

macerans H.-J.

Synthesis of I m m o b i l i z e d Peptide Fragments on Polystyrene-Polyoxyethylene for A f f i n i t y Chromatography ( I n German) 455

SCHEPERS, ST. B R I N G E R - M E Y E R , and H . S A H M

401 Semicontinuous and Continuous Production of Citric A c i d w i t h I m m o b i l i z e d Cells o f Aspergillus niger H . E I K M E I E R and H . J. R E H M

ABRAHAM,

P.

WASHAUSEN,

I . Z I E G L E R and ST. T H I E R F E L D E R

461

408

Microbial H y d r o x y l a t i o n o f Cedrol and Cedrene W.-R.

Biopterin Synthesis i n Mouse Spleen d u r i n g Bone M a r r o w Transplantation Correlates w i t h U n i m paired Hemopoietic Engraftment

and

KIESLICH

K. 414

In vivo Screening of Glutathione Related Detoxification Products in the Early State of D r u g D e v e l opment A.

P R O X , J.

S C H M I D , J.

NICKL,

and

G.

HARDT

6-Methylpurine, 6-Methyl-9-ß-D-ribofuranosylpurine, and 6 - H y d r o x y m e t h y l - 9 - ß - D - r i b o f u r a n o s y l purine as A n t i v i r a l Metabolites of Collybia maculata (Basidiomycetes) K . LEONHARDT, T. A N K E , E. H I L L E N - M A S K E ,

and

W . STEGLICH

420

Enzymatic Synthesis of Riboflavin and F M N Specifically Labeled w i t h C i n the Xylene R i n g 13

H . S E D L M A I E R , F. M Ü L L E R , P. J. K E L L E R , and BACHER

A.

ENGEL465

Synthesis and Complexing Features of an A r t i f i c i a l Receptor for Biogenic A m i n e s ( I n G e r m a n ) F. P. S C H M I D T C H E N

476

Metabolism o f the Herbicide 2-(2,4-Dichlorophenoxy)-propionic A c i d ( D i c h l o r p r o p ) i n Barley (Hordeum vulgare) G.

BÄRENWALD,

B.

SCHÜTTE

SCHNEIDER,

and

H.-R. 486

425

A V i t a m i n D Steroid H o r m o n e i n the Calcinogenic 3

Grass Trisetum

flavescens

W . A . R A M B E C K , H . W E I S E R , and H . Z U C K E R

Site Directed Antisera to the D - 2 Polypeptide Subunit o f Photosystem I I R. G E I G E R , R. J. B E R Z B O R N , B . D E P K A , W . O E T T -

430

MEIER, and A . T R E B S T

491

Contents

VII

Enzymatic A c y l o i n Condensation o f Acyclic A l d e -

Contents of Number 5

hydes Original

Communications

W.-R.

Epicuticular Wax Hydrocarbons o f Ericaceae in Germany

A B R A H A M and B . S T U M P F

Sn-Glycerol-3-phosphate

is a

559

Product

of

Starch

Degradation i n Isolated Chloroplasts from

I . SALASOO

499

mydomonas

Chla-

reinhardii

G . K L Ö C K and K . K R E U Z B E R G

O n the Essential Oils from Chrysanthemum

567

balsaSubstrate

mita L . ( I n German) H . STROBEL, K . K N O B L O C H , and E . Z I E G L E R

502

Flow

from

Photosynthetic

Carbon

Metabolism to Chloroplast Isoprenoid Synthesis in Spinach Evidence for a Plastidic Phosphoglyc-

Epoxy-rra/25-isodihydrorhodophytin, a N e w Metabolit from Laurencia S.

IMRE,

H.

D.

obtusa ( I n German)

LOTTER,

H.

WAGNER,

erate Mutase

and

R.

H.

THOMSON

507

SCHULZE-SIEBERT,

A.

HEINTZE,

and

G.

SCHULTZ

570

Energy-Dependent

Chlorophyll

Fluorescence

Quenching in Chloroplasts Correlated w i t h Q u a n Structure Elucidation o f K w a k h u r i n , a New Prenylated Isoflavone from Pueraria mirifica Roots S. T A H A R A , J . L . I N G H A M , and S. Z . D Z I E D Z I C

t u m Y i e l d o f Photosynthesis G . H . K R A U S E and H . L A A S C H

Spontaneous Biosynthesis of V i t e x i n and Isovitexin: Enzymatic Synthesis of the C-Glucosylflavones V i t e x i n and Isovitexin

with

Fagopyrum

an

Enzyme

esculentum

Preparation

in L - M S O Treated Chlorella

Malondialdehyde

from

S. A G A R W A L , A . G H O S H , and S. N . C H A T T E R J E E

585

519

Pyrimidine Homoribonucleosides: Synthesis, Solution C o n f o r m a t i o n , and Some Biological Properties P. L A S S O T A , J . T . K U S M I E R E K , R.

Cells

Y . S H I R A I W A and G . H . S C H M I D

525

of Arylsulfatase A c t i v i t y by Sulfate

STOLARSKI,

D . SHUGAR

fusca

I.

BIEDLINGMAIER,

and

SCHMIDT

and 589

Reactivation o f Streptolysin S by Oligonucleotide A . T A K E T O and Y . T A K E T O

Starvation in Chlorella S.

of

M . Seedlings

Effect o f p H on Glycolate and A m m o n i a Excretion

NIEDERMEYER,

Release

U l t r a v i o l e t Light Exposed L i p o s o m a l Membranes

from

F . KERSCHER and G . F R A N Z

Derepression

581

510

599

A. 530

R i n g O p e n i n g Reactions o f Bioreactive Lactam Systems ( I n German)

Root H a i r Specific Proteins in Glycine

H . FRISTER and E . S C H L I M M E

max 537

D . W E R N E R and A . B . W O L F F

Embryotoxicity

Induced

by

603

Alkylating

Agents.

Some Methodological Aspects o f D N A A l k y l a t i o n Synthesis,

Analysis and

Characterization

of

the

Coenzyme A Esters of o-Succinylbenzoic A c i d , an Intermediate in V i t a m i n K (Menaquinone) B i o 2

synthesis R . K O L K M A N N and E . L E I S T N E R

Studies in M u r i n e Embryos Using Ethylmethanesulfonate T.

PLATZEK,

G.

BOCHERT,

U.

RAHM,

and

NEUBERT

D. 613

542

Tetraponerine-8, an A l k a l o i d a l Contact Poison i n a Chloride A v a i l a b i l i t y Affects the Malate Content and its C o n t r o l by the Circadian Clock in Pulvini of Phaseolus coccineus L . W.-E. HAMPP

M A Y E R , W . A . R Ü G E , N . S T A R R A C H , and

R. 553

Neoguinean Pseudomyrmecine A n t ,

Tetraponera

sp. J . C. B R A E K M A N , D . D A L O Z E , J . M . P A S T E E L S , P. V A N H E C K E , J . P. D E C L E R C Q , V . S I N N W E L L , and

W.

FRANCKE

627

X

Contents Contents of N u m b e r

Original

Partial Characterization o f an Enzyme from the Fungus Ascochyta rabiei for the Reductive Cleavage of Pterocarpan Phytoalexins to 2'-Hydroxyisoflavans

7/8

Communications

Z Z E - C o n f i g u r a t i o n of C h r o m o p h o r e ß-153 in CPhycocyanin from Mastigoclaclus laminosus G.

S C H M I D T , S.

SIEBZEHNRÜBL,

R Ü D I G E R , H . SCHEER, T.

R.

FISCHER,

W.

SCHIRMER, W . B O D E ,

and

R . HUBER

845

B . H Ö H L and

W . BARZ

Comparing Short-Term Effects of A m m o n i a and Methylamine on Nitrogenase A c t i v i t y in Anabaena variabilis ( A T C C 29413) S. R E I C H , H . A L M O N , and

a-Diceroptene: A New D i m e r i c Structure for Isoceroptene C H . V I L A I N , A . HUBERT, L. DUPONT, K . R . MARKH A M , and

E . WOLLENWEBER

849

Stereochemistry o f T w o Hydroxybiflavanonols from Garcinia

cola Nuts

J. S O N N E N B I C H L E R , I . M A D U B U N Y I , and

H . SCHEER

855

897

P. B Ö G E R

902

Reversible pH-Induced Dissociation o f Glucose Dehydrogenase from Bacillus megaterium. I I . Kinetics and Mechanism E . M A U R E R and

G . PFLEIDERER

907

O n the Possible Relation between M o r p h o l o g y and Precursors of the Crystallities in Calcified Tissues F. C.

M . D R I E S S E N S , R.

A . TERPSTRA, P.

M A , J . H . M . W Ö L T G E N S , and

BENNE-

R. M . H . V E R B E E C K

916 "Epicuticular Waxes" from Exine Material of Pine Pollen C H . NIESTER, P . - G .

G Ü L Z , and

R. WIERMANN

858

In the Search for New Anticancer Drugs, X X I . Spin Labeled Nitrosoureas G . SOSNOVSKY, S. W . L I , and

Bryoflavone and Heterobryoflavone, T w o New Isoflavone-flavone Dimers f r o m Bryum capillare H . G E I G E R , W . S T E I N , R . M U E S , and

H . D.

ZINS-

MEISTER

863

Site of Lupanine and Sparteine Biosynthesis in Intact Plants and in vitro Organ Cultures M . WINK

868

Occurrence o f A m a v a d i n in Mushrooms Genus Amanita ( I n G e r m a n ) E . K O C H , H . K N E I F E L , and

of

the

E . BAYER

873

Ascorbic A c i d and Glutathione Contents of Spruce Needles from Different Locations in Bavaria W . F. O S S W A L D , H . SENGER, and

F. E . E L S T N E R

879 A c e t y l - C o A : 4 - H y d r o x y b u t i n y l b i t h i o p h e n e O-Acetyltransferase Isoenzymes f r o m Tagetes patula Seedlings G . M E T S C H U L A T and

R . SÜTFELD

A. SCHMIDT

921

Transmission of H o r m o n a l I m p r i n t i n g in Tetrahymena Cultures by Intercellular C o m m u n i c a t i o n G . C S A B A and

P. K O V A C S

932

M T D Calculations on Quantitative S t r u c t u r e - A c t i v i ty Relationships of Steroids Binding to the Progesterone Receptor M . B O H L , Z . S I M O N . A . V L A D , G. K A U F M A N N ,

and

K. PONSOLD

935

Synthesis of Heat Shock Proteins during A m i n o A c i d or Oxygen L i m i t a t i o n in Bacillussubtilis relA^ and relA ( I n German) M . H E C K E R , A . R I C H T E R , A . SCHROETER. L .

WÜL-

FEL, and F. M A C H

941

Sexual Pheromones and Related Egg Secretions in Laminariales (Phaeophyta) I . M A I E R , D . G. M Ü L L E R , G. L A N D , and

GASSMANN, W .

L . JAENICKE

BO948

885

Uptake and U t i l i z a t i o n of Sulfonic Acids in the Cyanobacterial Strains Anabaena variabilis and Plectonema 73110 S. B I E D L I N G M A I E R and

N. U. M . RAO

891

Comparison of D u f o u r Gland Secretions o f T w o Species of Leptothorax Ants ( H y m e n o p t e r a : Formicidae) M . F. A L I , E . D . M O R G A N , A . B . A T T Y G A L L E ,

and

J . P. J . B I L L E N

955

Contents Comparative

Sex Pheromone Biosynthesis

Obliquebanded ceana,

Leafroller,

in the

Choristoneura

rosa-

and the Redbanded Leafroller,

taenia velutinana,

XI

A r e Small R N A s Associated w i t h Crohn's Disease? R. P E C H A N , H . K U N E R T , and

H . J . GROSS

1006

ArgyroInhibition

Moths

S T . P. FOSTER and W . L . R O E L O F S

961

of c A M P Phosphodiesterase by Some

Phototherapeutic Agents L . B O V A L I N I , P. L U S I N I , S. S I M O N I , D . V E D A L D I , L .

Effect of H y d r o x y l a m i n e Derivatives on Photorespiration in the Tobacco tabacum G.

Aurea M u t a n t

A N D R E A S S I , F. D A L L ' A C Q U A , and P. M A R T E L L I

Nicotiana

1009

Su/su

H . S C H M I D , K . P.

BADER, A.

R A D U N Z , C.

V A N A S S C H E , N . R E I N I E R , and B . C O U R T I A D E

J.

Report

965

Chimeric Genes — T h e i r C o n t r i b u t i o n for Molecular Estimation o f the Extent of D N A Platination after Interaction of d s - D D P w i t h D N A and C h r o m a t i n I . K U L A M O W I C Z and Z . W A L T E R

Biology and Plant Breeding ( I n German) H . - H . STEINBISS and J . S C H E L L

1011

970

Reversed Light Reaction of the Screening Pigment in a C o m p o u n d Eye Induced by Noradrenaline A . J U S E , G . H O G L U N D , and K . H A M D O R F

Contents of Number

973

Light and E l e c t i o n Microscopic Studies Regarding

Original

Co mm

9/10

unications

Cell Contractility and Cell C o u p l i n g in L i g h t Sensitive Smooth Muscle Cells from the Isolated Frog Iris Sphincter 977

Fluorescence M o n i t o r i n g of Membrane Spatio-Temporal

Neurons of Helix

Resolution

Potentials: in

Isolated

pomatia 986

G r o w t h Kinetics of the G2-Phase of E h r l i c h Ascites T u r n o r Cells, Separated from Anaerobically T r e a t e d Asynchronous Cultures C. K R O L L . W . K R O L L , and F. S C H N E I D E R

991

of Statistically Significant Correla-

tions between 8 and 12 k H z Atmosferics and Sudden Deafness G.

RUHENSTROTH-BAUER, K.

P.

H A D J I E V A , S.

POPOV, B . B U D E V S K A , A .

DYUL-

GEROV, and S. A N D R E E V

1019

T h i n Layer Chromatographic and Infra Red Spectral Evidence for the Presence o f Phosphonolipids in

T H . K N Ö P F E L and P. F R O M H E R Z

Demonstration

Conopeum

sen rat urn

K. V . W O L F

the

Terpenoids from a Black Sea Bryozoan

Cicada

oni

M . C. M O S C H I D I S

1023

A d n e x a l Glands Chemistry o f Messor ebeninus (Formicidae: M y r m i c i n a e ) M . C O L L , A . H E F E T Z , and

Forel

H. A. LLOYD

1027

A N o v e l Caffeic A c i d D e r i v a t i v e and Other Constituents of Populus

B u d Excretion and Propolis

(Bee-Glue) M E E S , R.

SANDHA-

G E N , H . B A U M E R , and B . F I L I P I A K

999

E. WOLLENWEBER, Y . A S A K A W A , D . SCHILLO, U . L E H M A N N , and H . W E I G E L

Polyunsaturated

Pheromones:

(Z,Z)-6,9-Alkadienes

Notes

and

1030

Semi-Synthesis

of

( Z , Z , Z)-3,6,9-Alka-

trienes from N a t u r a l l y O c c u r r i n g Fatty Acids y-Decalactone, an Odoriferous C o m p o u n d from the M a l e Butterfly, Lethe marginalis Motschulsky N.

HAYASHI, H . KAWAGUCHI, A.

KOMAE

N I S H I , and

E.

SCHULTE

1035

H . 1001

The Phosphatidylinositol Species of Suspension C u l t u r e d Plant Cells S. H E I M and K . G . W A G N E R

H . K . M A N G O L D , H . B E C K E R , and

N M R Spectra o f Flavone Di-C-glycosides from Apometzgeria pubescens and the Detection of Rotational Isomerism i n 8-C-Hexosylflavones K . R. M A R K H A M , R.

1003

ZINSMEISTER

M U E S , M . S T O L L , and

H.

D. 1039

XII

Contents

Trichothecene Mycotoxins from Fusarium Cultures N.

C.

P.

BALDWIN,

B.

W.

culmorum

B Y C R O F T , P.

M.

D E W I C K , D . C. M A R S H , and J. G I L B E R T

1043

Comparison of Various Strategies Designed to Optimize Indole A l k a l o i d A c c u m u l a t i o n of a Cell Suspension Culture of Catharanthus roseus J. B E R L I N , C H . M O L L E N S C H O T T , and

F.

DICOSMO

1101 Metabolism o f the Prenylated Pterocarpan E d u n o l by Aspergillus flavus S. T A H A R A and J. L . I N G H A M

1050

A m p e r o m e t r i c T i t r a t i o n Largely Overestimates Chloride Concentrations in Chloroplast Extracts G . S C H R O P P E L - M E I E R and W . M . K A I S E R

Fungal Metabolism of the Prenylated Isoflavone 2,3Dehydrokievitone S.

T A H A R A , E.

M I S U M I , J.

MIZUTANI,

and

J.

INGHAM

L. 1055

1109

Extrachloroplastic Site o f Synthesis of Three C h l o r o plast Proteins in Maize (Zea mays) L.

A.

K L E C Z K O W S K I , C.

A.

Z E I H E R , and

D.

RANDALL

Novel Flavonoids from the Fern Notholaena phured

D. 1113

sulOrganolead Toxicity in Plants: T r i e t h y l Lead ( E t P b ) Acts as a Powerful Transmembrane C I " / O H ~ Exchanger Dissipating H - G r a d i e n t s at N a n o - M o l a r Levels +

F. J. A R R I A G A - G I N E R , M . I I N U M A , T . T A N A K A , M .

3

+

M I Z U N O , C. S C H E E L E , and E . W O L L E N W E B E R

1063

A . H A G E R , I . M O S E R , and

Synthesis of Piperoyl Coenzyme A Thioester U . SEMLER, G.

S C H M I D T B E R G , and

G.

G.

Geraniol-10-hydroxylase A c t i v i t y and Its Relation to Monoterpene Indole A l k a l o i d A c c u m u l a t i o n i n Cell Suspension Cultures o f Catharanthus roseus 0 . S C H I E L , L . W I T T E , and J. B E R L I N

1075

Effect o f p H on the Slow Phase Components o f Delayed Luminescence in Chloroplasts £ . H I D E G and

S. D E M E T E R

A New Detection Procedure for Aminoacylase A c tivity of Microorganisms D i r e c t l y on Plate Culture w i t h o-Phthalaldehyde ( I n German) KULA

1082 Small A n g l e X - R a y Study on the Structure of A c t i v e and Inactive Ribulose-l,5-bisphosphate Carboxylase-Oxygenase from Spinach. Evidence for a Configurational Change 1. P I L Z , E . S C H W A R Z , G . P. P A L , and

Quantitative Histochemical Analysis of Starch, Malate and K , together w i t h the A c t i v i t y o f Phospho-enolpyruvate Carboxylase along an Elongating Primary Leaf of Hordeum vulgare +

and

P. B R U N E R I E , I . B E N D A , G . B O C K , and

1126

In vitro T j and T Relaxation Times of Coagulating B l o o d and Thrombuses ( I n German) 2

U . M . L A N D L E R , K . H E R G A N , E . J U S T I C H , and STERK

H. 1135

Dependence of a Sleeping Parameter from the N-S or E - W Sleeping D i r e c t i o n G.

RUHENSTROTH-BAUER, E.

R Ü T H E R , and

REINERTSHOFER

TH. 1140

Notes A u t o m a t i c T u r g o r Pressure Recording in Plant Cells K . - H . B Ü C H N E R , G . W E H N E R , W . V I R S I K , and ZIMMERMANN

U. 1143

H . ZIEGLER

1092 Biotransformation of Citral by Botrytis

A . DE BOISFLEURY CHEVANCE

W . SAENGER

1089

R. H A M P P , W . H . O U T L A W Jr.,

1121

Chemokinesis and Necrotaxis o f H u m a n Granulocytes: the I m p o r t a n t Cellular Organelles H . G R U L E R and

M.-R.

1116

GROSS

1070

Y . Y A M A Z A K I , W . H U M M E L , and

W. BERTHOLD

cinerea

Distinct Substrate Specificity of D i h y d r o f l a v o n o l 4-Reductase from Flowers of Petunia hybrida G . F O R K M A N N and

B . RUHNAU

1146

P. SCHREIER

1097

Erratum

1149

Contents Contents of Number

11/12

XIII

4-(2'-Carboxyphenyl)-4-oxobutyryl Coenzyme A Ester, an Intermediate in V i t a m i n K (Menaquinone) Biosynthesis 2

Contents o f Nos 1-12

III-XIV

R. K O L K M A N N and

Original

SAUTER. E.

Betulin W.

H.

in

Archaeological

HAYEK,

W.

MOCHE,

U . JORDIS

Tar and 1151

Chemical Composition and M o r p h o l o g y o f E p i cuticular Waxes from Leaves o f Solanum tuberosum A . SEN 1153 A s y m m e t r i c Reduction of 4(5)-Oxocarboxylic Acids by Baker's Yeast ( I n G e r m a n ) M . G E S S N E R , C. G Ü N T H E R , and

A. MOSANDL

1159

Partial Lack of N - A c e t y l Substitution of G l u cosamine in the Peptidoglycan of the B u d d i n g Phototrophic Rhodomicrobium vannielii U.

J.

Dow,

1207

Communications

Identification of (In German) F.

E. LEISTNER

JÜRGENS, B.

RIETH, J.

W E C K E S S E R , C.

and W . A . K Ö N I G

S.

Role of Pyrophosphate: Fructose-6-phosphate 1-Phosphotransferase in Glycolysis in Cultured Catharanthus roseus Cells H . A S H I H A R A and T . H O R I K O S I

1215

The Separation o f T w o Different Enzymes Catalyzing the F o r m a t i o n of Hydroxycinnamic A c i d Glucosides and Esters P. A . B A U M K E R , M . J Ü T T E , and

R. W I E R M A N N

1223

Protein Sequence and Structure of N - T e r m i n a l A m i n o Acids of Subunit Delta of Spinach Photosynthetic ATP-Synthase CF, R.

J. BERZBORN, W.

F I N K E , J . O T T O , and

MEYER

H.

E. 1231

Isolation and Characterization of a Supramolecular Complex of Subunit I I I of the ATP-Synthase from Chloroplasts P. F R O M M E , E . J . B O E K E M A , and

P. G R Ä B E R

1239

1165

A c c u m u l a t i o n of Phenolic Compounds and Phytoalexins in Sliced and Elicitor-Treated Cotyledons o f Cicer arietinum L . U . J A Q U E S , H . K E S S M A N N , and W . B A R Z

1171

N a t u r a l Inhibitors of G e r m i n a t i o n and G r o w t h I V Compounds from Fruit and Seeds o f M o u n t a i n A s h (Sorbus aucuparia) U . O S T E R , I . B L O S , and W . R Ü D I G E R

1179

The Polyphasic Rise o f C h l o r o p h y l l Fluorescence upon Onset o f Strong Continuous I l l u m i n a t i o n : I . Saturation Characteristics and Partial C o n t r o l by the Photosystem I I Acceptor Side C H . N E U B A U E R and

U . SCHREIBER

1246

The Polyphasic Rise o f C h l o r o p h y l l Fluorescence upon Onset of Strong Continuous I l l u m i n a t i o n : I I . Partial C o n t r o l by the Photosystem I I D o n o r Side and Possible Ways o f Interpretation U . SCHREIBER and

C H . NEUBAUER

1255

The v versus v[I] Plot A . C. BORSTLAP

1185

Benzofuroxan as Electron Acceptor at Photosystem I B.

D e v e l o p m e n t of N e w Plate Tests for the Detection o f M i c r o b i a l Hydrolysis of Esters and Oxidations o f 2-Hydroxycarboxylic Acids ( I n German) Y . Y A M A Z A K I and

M.-R.

KULA

1187

E n z y m a t i c Synthesis o f 4 ' - and 3 ' , 4 ' - H y d r o x y l a t e d Flavanones and Flavones w i t h Flower Extracts o f Sinningia cardinalis K . S T I C H and G . F O R K M A N N

1193

Purification and Properties of Chalcone Synthase f r o m Cell Suspension Cultures of Soybean R. W E L L E and H . G R I S E B A C H

1200

LOTINA-HENNSEN,

A.

GARCIA,

M.

AGUILAR,

and M . A L B O R E S

1265

C A R S Investigation o f Changes in Chromophore Geometry of C-Phycocyanin from Mastigocladus laminosus Induced by T i t r a t i o n with p-Chloromercuribenzenesulfonate S. S C H N E I D E R , F. B A U M A N N , and U . K L Ü T E R

1269

I n h i b i t i o n by Sethoxydim o f Pigment A c c u m u l a t i o n and Fatty A c i d Biosynthesis in Chloroplasts o f Avena Seedlings H . K . L I C H T E N T H A L E R , K . K O B E K , and

K . ISHII

1275

Contents

XIV Bioenergetics Studies of the Cyanobacterium baena variabilis S. SCHERER, H . S A D O W S K I , and

Ana-

P. B Ö G E R

1280

Thermal Lability of Membrane Proteins of Age Separated Erythrocytes as Studied by Electron Spin Resonance Spin Label Technique G.

Radiochemical Methods for Studying Lipase-Catalyzed Interesterification of Lipids R. S C H U C H and

K. D. MUKHERJEE

1285

c A M P - D e p e n d e n t Protein Kinase A c t i v i t y in Yeast Mitochondria G . M Ü L L E R and

W . BANDI.OW

1291

Twisted Fibrils are a Structural Principle in the Assembly of Interstitial Collagens, Chordae Tendineae Included W.

FOLKHARD,

D.

E. K N Ö R Z E R . M .

CHRISTMANN, W . H.

J.

KOCH,

N E M E T S C H E K - G A N S L E R , and

E.

GEERCKEN. MOSLER,

T. NEMETSCHEK

Lysine Decarboxylase from Hafnia alvei: Purification, Molecular Data and Preparation o f Polyclonal Antibodies 1307

Recognition of H L A Class I I Molecules by A n t i p e p tide Antibodies Elicited by Synthetic Peptides Selected from Regions of H L A - D P Antigens A. and

C H E R S I , R.

A.

HOUGHTEN, M .

C.

MORGANTI.

E. M U R A T T I

BARTOSZ, G.

CHRIST, H .

1313

1343

Lipids in the Gular Gland Secretion of the A m e r i c a n A l l i g a t o r (Alligator mississippiensis) P. J . W E L D O N , A . S H A F A G A T I , and

K . W I T T H O H N and

and

J. W . WHEELER

1345 Identification of the Sex Pheromone of Eggplant Borer Leucinodes orbonalis Guenee ( L e p i d o p tera: Pyralidae) ZHU Yu

PINGCHOU, YONGQING,

KONG

FANLEI,

YU

J I N SHUPING, H U

( Z ) - 5 - D o d e c e n - l - o l . A n o t h e r I n h i b i t o r o f Pheromonal A t t r a c t i o n in Coleophora laricella E . PRIESNER

1349

(Z)-3-TetradecenyI Acetate as a Component in Gelechiinae and (Lepidoptera: Gelechiidae)

Sex-Attractant Anomologinae

E . PRIESNER

1352

M a m m a l i a n Pheromone Studies. V I . Compounds from the Preorbital Gland of the Blue D u i k e r , Cep halophus mon tico I a P. J . PRETORIUS

1319

SHANI.

S.

BENITA,

M.

ABDULRAZIK,

and 1323

M H C - A n t i g e n s : Constituents of the Envelopes o f H u m a n and Simian Immunodeficiency Viruses R.

GELDERBLOM. K U N Z E , and

H.

REUPKE.

T.

WINKEL.

G . PAULI

Evidences for O r c a d i a n Mutant of Drosophila

1328

Rhythmicity in the melanogaster

C H . H E L F R I C H and W . E N G E L M A N N

per° 1335

Conformational Changes in Proteins Induced L o w Temperatures: an Infrared Study H. L. CASAL, U . KÖHLER, H . H .

Inhibition of the A c e t y l - C o A Carboxylase of Barley Chloroplasts by Cycloxydim and Sethoxydim M . FOCKE and

H . K . LICHTENTHALER

1361

Formation of Large Thioredoxin / Accompanies Chloroplast Development in Scenedesmus obliquus P. L A N G L O T Z and

H. FOLLMANN

1364

Cellular Spin Resonance of Yeast in a Frequency Range up to 140 M H z R. H O L Z E L and

I . LAMPRECHT

1367

M e l t i n g Pressure. V o l u m e and Stability of B l o o d at H i g h Pressure ( I n German)

Notes

G O N I , and J. L . R. A R R O N D O

1358

Radioprotective

A. YERUSHALMI

H.

1355

Structural Features and Biological Functions in Blue Copper Proteins Y. NISHIDA

J.

and 1347

Other

C. M . N A U M A N N

Efficacy of Sustained-Release Drugs in vivo

SHENGDI,

XINHUA,

Yu JIANWEI

B . V . BURGER and

Active Cyanogenesis — in Zygaenids Lepidoptera

BOSSE. R. S T E P H A N ,

H . GÄRTNER

H. 1303

H . B E I E R , L . F. FECKER, and J. B E R L I N

and

M A N T S C H , F.

by M. 1339

A . K L U G E and

H. LENTZ

1370

Subjekt Index

1373

A u t h o r s Index

1403

Chromophore Assignment in C-Phycocyanin from Mastigocladus laminosus S. S i e b z e h n r ü b l , R . Fischer, and H . Scheer Botanisches Institut der Universität München, Menzinger Straße 47, D-8000 München 19, Bundesrepublik Deutschland Z. Naturforsch. 42c, 258-262 (1987); received November 4, 1986 Photosynthesis, Cyanobacteria, Energy Transfer, Antenna Pigments, Cystein Modification C-phycocyanin from the cyanobacterium, Mastigocladus laminosus, and its subunits have been treated with p-chloromercuribenzenesulfonate (PCMS). A single reactive site was found on the ßsubunit, and assigned to the single free cystein-ßl09. The concomitant spectral changes (absorption, fluorescence, circular dichroism), together with the known close proximity of cys-ßl09 to chromophore ß82, allowed an unambiguous assignment of the three spectrally, biochemically and functionally different chromophores to specific binding sites on the two peptide chains (a84: 616-618, ß82: 622-624, ß 153: 598-600 nm).

Introduction C-phycocyanins ( P C ) , the light-harvesting pigments from cyanobacteria, are composed o f t w o subunits ( a , ß) bearing one and t w o open-chain tetrapyrrolic chromophores, respectively [1—5]. I n spite o f their common structures [ 1 , 2, 6, 7 ] , the three chromophores differ in their spectroscopic properties [8—10] and their reactivities [ 1 0 - 1 2 ] . These differences, which arise from the different environments of the individual chromophores in the native chromoprotein, are essential for the fine tuning of their biological functions [1—5, 8, 10, 13]. The individual environments and conformations o f the three chromophores, have recently been mapped in detail by sequenation [14—19] and X-ray crystallography [20, 21]. The data indicate a considerable degree of homology around the respective chromophores in PC's from different organisms. The single chromophore on the a-subunit is attached to Cys-84, and the chromophores on the ß - s u b u n i t to Cys-82 and Cys-153. The t w o PC's studied by high-resolution X-ray crystallography [20, 21] (from Mastigocladus laminosus and Agmenellum quadruplicatum), have furthermore almost identical chromophore and protein conformations and differ only i n their aggregation state. A correlation between the spectrally and structurally distinct chromophores has hitherto been lacking, but is very important i n view of the recent progress i n energy transfer studies (see e.g. [13]). Based on the Reprint requests to Prof. Dr. H . Scheer. Verlag der Zeitschrift für Naturforschung, D-7400 Tübingen 0341-0382/87/0300-0258 $ 01.30/0

Observation o f a single binding site ( c y s - ß l 0 9 ) for mercurials [20, 21], we have now titrated PC and its subunits from M. laminosus with the t h i o l reagent, P C M S , which led to the unambiguous assignment of the chromophores. This situation is more clear-cut than in other biliproteins, e.g. p h y c o e r y t h r i n [22] containing more than a single free cystein available for reaction w i t h mercurials [ 1 - 5 ] .

Materials and Methods M. laminosus was grown in Castenholz m e d i u m [23] in 300 1 batch cultures [24] and p r o v i d e d to us and stored deep frozen. PC and the subunits were prepared as described previously [ 6 ] . p - C h l o r o - m e r curi-benzenesulfonic acid ( P C M S , A l d r i c h ) was a gift of T . Schirmer, all other chemicals came f r o m M e r c k (Darmstadt). A b s o r p t i o n spectra were recorded on a model 8 4 5 1 A spectrophotometer (Hewlett-Packard), fluorescence spectra on a model D M R 2 2 fluorimeter (Zeiss, Oberkochen) and circular dichroism spectra on a model V dichrograph ( J o b i n - Y v o n , U n t e r haching). Titrations were carried out by adding aliquots o f a stock solution o f P C M S (2.26 I H M , i n potasium phosphate buffer, 100 ITIM, p H 7.0) to the c h r o m o p r o t e i n solutions (1—3 \XM i n the same buffer). The following extinction coefficients were used for the concentrat i o n determinations: PC trimer = 870,000 c m V corresponding to 290,000 per p r o t o m e r (aß)\ and 97,000 per c h r o m o p h o r e ) ; ß - s u b u n i t = 164,000 corresponding to 82,000 per chromophore; a-subunit = 122,000. They were determined by unfolding 1

S. Siebzehnrübl et al. • Chromophore Assignment in C-Phycocyanin from Mastigocladus laminosus the proteins w i t h 8 M urea, on the basis o f an extinction coefficient o f 15,400 per chromophore i n the denatured state [11].

Results Integral

PC

Treatment o f PC t r i m e r w i t h a threefold molar excess o f P C M S ( o n a monomer basis for the protein) results in a partial bleaching of the major absorption (612 n m ) (Fig. l a ) . I t is accompanied by a

300

400

500

600

m i n o r blue-shift and the f o r m a t i o n of a longerwavelength shoulder. The integrated negative band (612 n m ) i n the difference spectrum, is about twice as large as the concomitant increase (655 n m ) . There is also a distinct increase i n the near-uv band. T i t r a t i o n o f PC gives a saturation o f the reaction at 1 m o l P C M S per mole PC ( F i g . 2 a). The same general features are observed i n PC t r i m e r containing linker peptides ( X = 629 n m ) , but here the negative peak in the difference spectrum is located at 632 n m (data not shown). m a x

700

X (nm)

300

400

500

600 X (nm)

259

700

S. Siebzehnrübl etal • Chromophore Assignment in C-Phycocyanin from Mastigocladus laminosus

260

AA 0.08

- 0.04

0.00

-0.04

0.08

400

500

600 X(nm)

IC Fig. 1. Treatment of integral PC-trimer ( A ) , its ß-subunit (B) and its a-subunit (C) with PCMS. Absorption (left scales) and absorption difference spectra (right scales). 1 A : PC (4.8 nmol, spectrum a) treated with 13.6 nmol PCMS. Difference spectra recorded after 30 (c), 60 (d) and 100 min (e), final absorption spectrum (b) 180 min after addition. I B : Absorption spectrum (a) of the ßsubunit (5.66 nmol) and final difference spectrum (b) after addition of PCMS (9.06 nmol, / = 1 8 m i n ) . 1C: Absorption spectrum (a) of the a-subunit (6.9 nmol) and final difference spectrum (b) after addition of PCMS (9.1 nmol, / = 10 min).

2B

660

-650

Fig. 2. Titration of integral PC and its ß-subunit with PCMS. 2 A : Amplitude of the absorption difference signal (maximum-to-minimum). The absorptions before addition of the reagent were 0.68 (—-), 0.62 ( ) and 0.66 (•••). Labels see inset. 2B: Relative fluorescence emission intensities ( I , left scale) and maxima ( X in nm, right scale). Labels see inset. F

max

3

4 PCMS / Pigment

S. Siebzehnrübl et ai • Chromophore Assignment in C-Phycocyanin from Mastigocladus laminosus The fluorescence of PC trimer is reduced to a m i n i m u m of 3 5 % o f the original value and saturates at « .8 m o l P C M S , its wavelength is shifted by 10 nm to the red ( F i g . 2 b ) . I n the C D spectrum, the most notable feature is the development o f a shoulder on the red-wing o f the long-wavelength band at 650 n m (data not shown, similar to the ß - s u b u n i t shown in Fig. 3 ) . Subunits The spectral changes of the ß - s u b u n i t are very similar to the ones observed in trimeric PC. The absorption difference is again centered to the red of the absorption band. The amplitude o f the difference spectrum ( m i n i m u m to m a x i m u m ) is even larger than PC i n integral ( F i g . l b ) , and the reaction saturates at « . 6 5 m o l P C M S (Fig. 2). The fluorescence decreases by 4 5 % , and the amplitude of the red C D m a x i m u m by 7 % w i t h a concomitant rise of a shoulder around 650 n m and an increase of the near-uv band by 17% ( F i g . 3 ) . The a-subunit is essentially unreactive w i t h P C M S ( F i g . 1 c). There is only a minor feature in the difference spectrum w i t h an intensity of 6% as compared to the ß - s u b u n i t , which probably corresponds to a m i n o r contamination w i t h the latter ( « 5 % according to S D S - P A G E ) . The changes i n the fluorescence and the C D spectra are negligible as well (data not shown).

261

Discussion Besides the three cystein residues bound to the chromophores, PC contains only a single free cystein at position 109 on the ß - c h a i n . I n the native c h r o m o p r o t e i n , this cystein is very close ( ~ 4 A ) to the ß 8 2 c h r o m o p h o r e , w i t h an essentially proteinfree hole between t h e m , which is the single binding site for mercurials used for heavy-atom substitution [20, 21]. The distances to the other chromophores ( a 84, ß 153) b o t h on the same m o n o m e r and on the other monomers i n the trimer are considerably larger ( > 2 2 A ) . I t is, therefore, expected that the ß 8 2 chromophore is affected rather specifically upon binding o f mercurials. A differential response o f i n dividual chromophores has earlier been reported for phycoerythrin [22], but could not be explored further due to lack o f structural data at that t i m e . The saturation behavior i n solution confirms the binding o f only a single molecule of P C M S both i n the PC p r o t o m e r and its ß - s u b u n i t . This is further corroborated by the lack of reaction w i t h the a-subunit bearing no free cystein. A l l spectra can be rationalized by a single chromophore absorbing around 620 n m i n the ß - s u b u n i t and the t r i m e r , and around 630 n m in the linker-containing t r i m e r , respectively, being affected by the reaction of the free cys-109 w i t h P C M S . T w o spectrally [10, 13], chemically [10, 11] and photochemically [12] distinct chromophores have been identified on the ß - s u b u n i t

2.00QE-4

en in cci ac Q

-1.600E250.0

Fig. 3. Circular dichroism spectra of the ß-subunit before (a) and after (b) reaction with PCMS. The spectra have been taken from the samples shown in Fig. 1B in absorption (traces a and b).

262

S. Siebzehnrübl et al. • Chromophore Assignment in C-Phycocyanin from Mastigocladus laminosus

of PC from M. laminosus. One of them ( ß l ) absorbs around 620 n m , is only weakly optically active and strongly fluorescent ( i n the absence o f any acceptors), and the reversible photochemistry observed i n partly denatured PC or its ß - s u b u n i t resides on i t . The other one ( ß 2 ) absorbs below 600 n m , is strongly optically active and weakly fluorescent due to efficient energy transfer to ß 1 and (in integral PC) to the a-chromophore(s), and is photochemically inactive. I t is clear f r o m the data, that the former chromophore ( ß l ) is the one close to the P C M S binding site and hence identical w i t h ß 8 2 , whereas the latter ( ß 2 ) is identical w i t h chromophore ß 153. I f judged f r o m the absorption changes (increase in the near-uv, decrease in the visible band), the ß 8 2 chromophore assumes a more helical conformation [11] after PCMS b i n d i n g . This assignment relates the photochemical reactivity to the chromophore, ß 8 2 , which is least well defined in the X - r a y structure [20, 21] and hence probably also least rigidly bound. The data obtained w i t h the linker-containing t r i m e r ( X = 629 nm) also i n dicate, that the ß 8 2 chromophore is the one shifted towards longer wavelengths (from «622 to « 6 3 2 n m ) . These linkers, which are substantial in the spectral modulation and organisation of biliprotein aggregates, are most likely located in the inner m a x

[1] W. Rüdiger, Ber. Deutsch. Bot. Ges. 92, 413 (1979). [2] E . Gantt, Ann. Rev. Plant Physiol. 32, 327 (1981). [3] H . Scheer, in: Light Reaction Path of Photosynthesis (F. K. Fong, ed.), pp. 7-45, Springer Verlag, Berlin 1983. [4] A. N. Glazer, Ann. Rev. Biochemistry 52, 125 (1983). [5] W. Wehrmeyer, in: Proteins and nucleic Acids in Plant Systematics (U. Jensen and D. E . Fairbrother, eds.), Springer Verlag, Berlin 1983. [6] W. Kufer, O. Schmid, G . Schmidt, and H. Scheer, Z. Naturforsch. 41c, 437 (1986). [7] J . C . Lagarias, A. N. Glazer, and H . Rapoport, J . Am. Chem. Soc. 101, 5030 (1979); see however: H . Rapoport, and A . N. Glazer, in: Optical properties and structure of Tetrapyrroles (G. Blauer and H . Sund, eds.), pp. 411-423, de Gruyter, Berlin, New York 1985. [8] F . W. J . Teale and R. E . Dale, Biochem. J . 116, 161 (1970). [9] J . Friedrich, H . Scheer, B. Zickendraht-Wendelstadt, and D. Haarer, J. Amer. Chem. Soc. 101, 1030 (1981). [10] M. Mimuro, P. Füglistaller, R. Rümbeli, and H . Zuber, Biochim. Biophys. Acta 848, 155 (1986). [11] W. Kufer and H . Scheer, Hoppe-Seyler's Z. Physiol. Chem. 360, 935 (1979). [12] W. John, R. Fischer, S. Siebzehnrübl, and H . Scheer, in: Antennas and Reaction Centers of Photosynthetic Bacteria: Structure, Interaction and Dynamics (M. E . Michel-Beyerle, ed.), Springer Verlag, Berlin 1985.

hole of the doughnut-shaped t r i - and hexamers [ 4 , 5 ] . I n PC, the ß 8 2 chromophore partly extends i n t o this hole, which nicely fits w i t h its spectral shift u p o n binding o f linker peptides. Treatment w i t h the m e r c u r i a l , P C M S , thus allows a straightforward chromophore assignment i n PC and its aggregates. I t should be useful as well w i t h other biliproteins i n view of the increased structural data available [1—5]. Interestingly, there are also large differences among the binding kinetics of P C M S to the different PC aggregates including phycobilisomes. This may be helpful i n topological studies and is currently explored. Acknowledgements This w o r k was supported by the Deutsche Forschungsgemeinschaft, B o n n . W e are grateful for many stimulating discussions w i t h and the provision of unpublished material by T . Schirmer and R . H u b e r ( M a r t i n s r i e d ) . The large-scale culture of M. laminosus by G . Reng at the Gesellschaft für biotechnologische Forschung ( S t ö c k h e i m ) is acknowledged. Note added in proof: The amino acid residue numbering of the ß-subunit has been used differently in the literature [14, 20]. The one used corresponds to the original sequence data [14]. [13] K. Sauer, H . Scheer, and P. Sauer, Photochem. Photobiol., submitted; in part published in Proc. VII Int. Congr. Photosynthesis (J. Biggins, ed.), p. I. 1. 139, Martinus Nijhoff, Dordrecht. [14] G . Frank, W. Sidler, H . Widmer, and H . Zuber, H. Hoppe-Seyler's Z . Physiol. Chem. 359, 1491 (1978). [15] P. Freidenreich, G. S. Apell, and A . N. Glazer, J . Biol. Chem. 253, 212 (1978). [16] G . D. Offner, A. S. Brown-Mason, M. M. Ehrhardt, and R. F . Troxler, J . Biol. Chem. 256, 2167 (1981). [17] R. F . Troxler, M. M. Ehrhardt, A. S. Brown-Mason, and G . D. Offner, J. Biol. Chem. 256, 2176 (1981). [18] T. J . Pilot and J. L . Fox, Proc. Natl. Acad. Sei. USA 81, 6983 (1984). [19] R. Lorimier, D. A. de Bryant, R. D. Porter, W. Y . Liu. E . Jay, and S. E . Stevens jr., Proc. Natl. Acad. Sei. U S A 81, 7946 (1984). [20] T. Schirmer, W. Bode, R. Huber, W. Sidler. and H . Zuber, J. Mol. Biol. 184, 257 (1985). [21] T. Schirmer, R. Huber, M. Schneider, W. Bode, M. Miller, and M. L . Hackert, J . Mol. Biol. 188, 651 (1986). [22] J . Pecci, and E . Fujimori, Biochem. Biophys. Acta 154, 332 (1968). [23] R. W. Castenholz, Schweizer Z. Hydrol. 35, 538 (1970). [24] 300 1 mass cultures in the phototrophic fermenter at the Gesellschaft für Biotechnologische Forschung, 3301 Stöckheim, F R G .