by Î. Robert, A. Vermeglio, R. Steiner, Î. Scheer and Î. Lutz. 355 ... by R. van Grondelle, H. Bergström, V. Sundström, R.J. van Dorssen,. M. Vos and C.N. ...
Photosynthetic Light-Harvesting Systems Organization and Function
Proceedings of an International Workshop October 12-16,1987 Freising, Fed Rep. of Germany Editors Hugo Scheer · Siegfried Schneider
W G Walter de Gruyter · Berlin · New York 1988 DE
CONTENTS
L i s t of Participants SECTION I .
XIII
ORGANIZATION: BIOCHEMICAL METHODS
Introduction: The Biochemistry of Light-Harvesting Complexes by R.J. Cogdell
1
Phycobilisome-Thylakoid I n t e r a c t i o n : The Nature of High Molecular Weight Polypeptides by E. Gantt C.A. Lipschultz and F.X. Cunningham Jr
11
On the Structure of Photosystem II-Phycobilisome Complexes of Cyanobacteria by E. Mörschel and G.-H. Schatz
21
Structure of Cryptophyte Photosynthetic Membranes by W. Wehrmeyer
35
Structural and Phylogenetic Relationships of Phycoerythrins from Cyanobacteria, Red Algae and Cryptophyceae by W. Sidler and H. Zuber
49
I s o l a t i o n and Characterization of the Components of the Phycobilisome from Mastigocladus laminosus and Crossl i n k i n g Experiments by R. Rümbeli and H. Zuber
61
C-Phycocyanin from Mastigocladus laminosus: Chromophore Assignment i n Higher Aggregates by Cystein Modification by R. Fischer, S. Siebzehnrlibl and H. Scheer
71
Photochromic Properties of C-Phycocyanin by G. Schmidt, S. Siebzehnrübl, R. Fischer and H. Scheer
77
Concerning the Relationship of Light Harvesting B i l i proteins t o Phycochromes i n Cyanobacteria by W. Kufer
89
Subunit Structure and Reassembly of the Light-Harvesting Complex from Rhodospiri Hum rubrum G9+ by R. Ghosh, ThT~Rosatzin ancTR. Baclofen
93
Primary Structure Analyses of Bacterial Antenna Polypeptides - Correlation of Aromatic Amino Acids with Spectral Properties - Structural S i m i l a r i t i e s with Reaction Center Polypeptides by R.A. Brunisholz and H. Zuber
103
VIII 1
The Structure o f the "Core * of the Purple Bacterial Photo synthetic Unit by D.J. Dawkins, L.A. Ferguson and R.J. Cogdell
115
A Comparison o f the Bacteriochlorophyl1 C--Binding Proteins of Chlorobium and Chloroflexus by P.D. Gerola, P. Hpjrup and J.M. Olson
129
I n t e r a c t i o n s between Bacteriochlorophyl1 c Molecules i n Oligomers and i n Chlorosomes of Green Photosynthetic Bacteria by D.C. Brune, G.H. King and R.E. Blankenship
141
Light-Harvesting Complexes of Chlorophyll c-Containing Algae by A.W.D. Larkum and R.G. H i l l e r
153
I s o l a t i o n and Characterization of a Chlorophyll a/c-Heteroxanthin/Diadinoxanthin Light-Harvesting Complex from Pleurochloris meiringensis (Xanthophyceae) by C. Wilhelm, C. Bliche I and B. Rousseau
167
The Antenna Components of Photosystem I I with Emphasis on the Major Pigment-Protein, LHC l i b by G.F. Peter and P. Thornber
175
SECTION I I :
ORGANIZATION: MOLECULAR GENETICS AND CRYSTALLOGRAPHY
Molecular Biology of Antennas by G. Drews
187
High-Resolution Crystal Structure of C-Phycocyanin and Polarized Optical Spectra of Single Crystals by T. Schirmer, W. Bode and R. Huber
195
C r y s t a l l i z a t i o n and Spectroscopic Investigation of Purple Bacterial B800-850 and RC-B875 Complexes by W. Welte, Τ. Wacker and A. Becker
201
Structure of the Light-Harvesting Chlorophyll Complex from Chloroplast Membranes by W. Kühlbrandt
211
a/b-Protein
PhycobiIi somes of Synchechococcus Sp. PCC 7002, Pseudanabaena Sp. PCC 7409, and Cyanopnora paracioxa: An "Analysis by" HoTecular Genetics by D.A. Bryant
217
Organization and Assembly of Bacterial Antenna Complexes by G. Drews
233
IX The Use of Mutants t o Investigate the Organization of the Photosynthetic Apparatus of Rhodobacter sphaeroides by C.N. Hunter and R. van Grondel le 77.77
247
Mechanisms o f P l a s t i d and Nuclear Gene Expression During Thylakoid Membrane Biogenesis in Higher Plants by P. Westhoff, Η. Grüne, Η. Schrubar, Α. Oswald, Μ. Streubel, U. Ljungberg and R.G. Herrmann
261
SECTION I I I :
ORGANIZATION: SPECIAL SPECTROSCOPY TECHNIQUES AND MODELS
Assigment o f Spectral Forms i n the Photosynthetic Antennas to Chemically Defined Chromophores by A. Scherz
277
Linear Dichroism and Orientation of Pigments in Phycobilisomes and t h e i r Subunits by L. Juszcak, N.E. Geacintov, B.A. Zilinskas and J. Breton
281
Low Temperature Spectroscopy of Cyanobacteria! Antenna Pigments by W. Köhler, J . F r i e d r i c h , R. Fischer and H. Scheer
293
Chromophore Conformations i n Phycocyanin and Allophycocyanin as Studied by Resonance Raman Spectroscopy by B. Szalontai, V. Csizmadia, Z. Gombos, K. Csatorday and M. Lutz
307
Coherent Anti-Stokes Raman Spectroscopy of Phycobi1isomes, Phycocyanin and Allophycocyanin from Mastigocladus laminosus by S. Schneider, F. Baumann, W. Steiner, R. Fischer, S. Siebzehnrlibl and H. Scheer
317
Optical Absorption and Circular Dichroism of Bacteriochlorophyl1 Oligomers i n Triton X-100 and i n the Light-Harvesting-Complex B850; A Comparative Study by V. Rozenbach-Belkin, P. Braun, P. Kovatch and A.Scherz
323
Absorption Detected Magnetic Resonance i n Zero Magnetic Field on Antenna Complexes from Rps. acidophila 7050 - The Temperature Dependence of tfie CarotenoTiTTrTpTet State Properties by J . U l l r i c h , J.U. v. Schütz and H.C. Wolf
339
Effect of Lithium Dodecyl Sulfate on Β 800-850 Antenna Complexes from Rhodopseudomonas acidophila: A Resonance Raman Study by B. Robert and H. Frank
349
χ
Bacteriochlorophyl 1 a/b i n Antenna Complexes of Purple Bacteria by Β. Robert, A. Vermeglio, R. Steiner, Η. Scheer and Μ. Lutz
355
Bacteriochlorophyll c Aggregates i n Carbon Tetrachloride as Models f o r Chlorophyll Organization i n Green Photo synthetic Bacteria by J.M. Olson and J.P. Pedersen
365
Orientation o f the Pigments i n the Reaction Center and the Core Antenna o f Photosystem I I by J. Breton, J . Duranton and K. Satoh
375
Non-Linear Absorption Spectroscopy of Antenna Chlorophyll a in Higher Plants by D. Leupold, H. S t i e l and P. Hoffmann
387
SECTION IV:
FUNCTION: ELECTRONIC EXCITATION AND ENERGY TRANSFER
Excitation Energy Transfer i n Photosynthesis by R. van Grondelle and V. Sundström
403
Fluorescence Spectroscopy of Allophycocyanin Complexes from Synechococcus 6301 Strain AN112 by P.Maxson, KTTauer and A.N. Glazer
439
Picosecond Energy Transfer Kinetics i n Allophycocyanin Aggregates from Mastigocladus laminosus by E. Bittersmann, W. Reuter, W. Wehrmeyer and A.R. Holzwarth
451
Picosecond Time-Resolved Energy Transfer Kinetics w i t h i n C-Phycocyanin and Allophycocyanin Aggregates by T. G i l l b r o , A. Sandström, V. Sundström, R. Fischer and H. Scheer
457
Energy Transfer i n "Native" and Chemically Modified C-Phycocyanin Trimers and the Constituent Subunits by S. Schneider, P. Geiselhart, F. Baumann, S. Siebzehnrübl, R. Fischer and Η. Scheer
469
Effect of Protein Environment and Excitonic Coupling on the Excited-State Properties of the Bilinchromophores i n C-Phycocyanin by S. Schneider, Ch. Scharnagl, M. Dürring, T. Schirmer and W. Bode
483
Excitation Energy Migration i n C-Phycocyanin Aggregates Isolated from Phormidium luridum: Predictions from the Förster's Inductive Resonance Theory by J . Grabowski and G.S. Björn
491
XI
Energy Transfer Calculations f o r two C-Phycocyanins Based on Refined X-Ray Crystal Structure Coordinates of Chromophores by K. Sauer and H. Scheer
507
Energy Transfer in Light-Harvesting Antenna of Purple Bacteria Studied by Picosecond Spectroscopy by V. Sundström, H. Bergström, Τ. G i l l b r o , R. van Grondelle, W. Westerhuis, R.A. Niederman and R.J. Cogdell
513
Excitation Energy Transfer i n the Light-Harvesting Antenna of Photosynthetic Purple Bacteria: The Role of the Long-WaveLength Absorbing Pigment B896 by R. van Grondelle, H. Bergström, V. Sundström, R.J. van Dorssen, M. Vos and C.N. Hunter
519
The Function of Chlorosomes i n Energy Transfer in Green Photosynthetic Bacteria by R.J. van Dorssen, M. Vos and J . Amesz
531
Energy Transfer in Chloroflexus aurantiacus: Effects of Temperature and TSaerobic^onTirfons by B.P. Wittmershaus, D.C. Brune and R.E. Blankenship
543
I n t e r p r e t a t i o n of Optical Spectra of Bacteriochlorophyl 1 Antenna Complexes by R.M. Pearlstein
555
Time Resolution and Kinetics of "F680" at Low Temperatures i n Spinach Chloroplasts by R. Knox and S. Lin
567
Picosecond Studies of Fluorescence and Absorbance Changes i n Photosystem I I Particles from Synechococcus Sp. by A.R. Holzwarth, G.H. Schatz and H. Brock
579
Analysis of Excitation Energy Transfer i n Thylakoid Membranes by the Time-Resolved Fluorescence Spectra by M. Mimuro
589
V. CONCLUDING REMARKS Future Problems on Antenna Systems and Summary Remarks by E. Gantt
601
Author Index
605
Subject Index
609
COHERENT ΑΝΤΙ-STOKES RAMAN SPECTROSCOPY OF
PHYCOBILISOMES,
PHYCOCYANIN AND ALLOPHYCOCYANIN FROM MASTIGOCLADUS LAMINOSUS
S. S c h n e i d e r ,
F. Baumann, W. S t e i n e r
I n s t i t u t für P h y s i k a l i s c h e und T h e o r e t i s c h e Chemie d e r TU-München, L i c h t e n b e r g s t r . 4, D-8046 G a r c h i n g R. F i s c h e r , S. Siebzehnrübl, H.
Scheer
B o t a n i s c h e s I n s t i t u t d e r Ludwig-Maximilians-Universität M e n z i n g e r s t r . 67, D-8000 München 19
Introduction I n order t o p r o v i d e a l a r g e g e o m e t r i c a l and s p e c t r a l
cross-
s e c t i o n f o r the a b s o r p t i o n of l i g h t , r e d and b l u e - g r e e n produce t h e s o - c a l l e d p h y c o b i l i s o m e s the chromoproteins p h y c o e r y t h r i n phycocyanin
(or p h y c o e r y t h r o c y a n i n ) ,
(PC) and a l l o p h y c o c y a n i n
(APC). Although PC and
APC c o n t a i n the same chromophore, p h y c o c y a n o b i l i n , p r o p e r t i e s of t h e s e b i l i p r o t e i n s bound to a p o p r o t e i n ) energies a r e modified
algae
(PBS), which c o n t a i n
the s p e c t r a l
( t e t r a p y r r o l chromophore
are quite d i f f e r e n t . Their e x c i t a t i o n a c c o r d i n g to t h e i r s p e c i a l f u n c t i o n i n
the l i g h t h a r v e s t i n g system
( s e e e.g. 1,2) by chromophore-
protein interactions (14). Resonance-enhanced Coherent A n t i - S t o k e s Raman
Spectroscopy
(CARS) has proved t o be a v e r y s u i t a b l e technique
to produce
v i b r a t i o n a l s p e c t r a of h i g h l y f l u o r e s c i n g chromophores as e.g.
l i g h t h a r v e s t i n g pigments
(3,4,5). I n this contribution
CARS-spectra of room-temperature s o l u t i o n s of and of phycocyanin c l a d u s laminosus
phycobilisomes
and a l l o p h y c o c y a n i n t r i m e r s from Mastigo-
a r e p r e s e n t e d and the i m p l i c a t i o n s to chromo-
phore s t r u c t u r e a r e d i s c u s s e d .
Photosynthetic Light-Harvesting Systems © 1988 Walter de Gruyter & Co., Berlin · New York
318
Experimental Resonance-enhanced CARS s p e c t r a a r e o b t a i n e d by f o c u s s i n g two l a s e r beams w i t h d i f f e r e n t wavelengths
λ
pump
(fixed)
and
( v a r i a b l e ) i n the sample, where by a n o n - l i n e a r e f f e c t
X s t o k e s
the CARS beam i s generated, VCARS
=
Vpuip
VCARS
>Vpuap
+
(Vpump
>Vstokes
which h o l d s the e q u a l i t y
- V s t o k e s )
, with
V=
1/X
1
[cm- ]
A p l o t of the CARS i n t e n s i t y as f u n c t i o n of r e v e a l s a v i b r a t i o n a l spectrum
Vpuap
-
Vstokes
of the chromophore. As
CARS frequency i s h i g h e r than e i t h e r pump and Stokes
the frequency,
no f l o u r e s c e n c e problems as i n spontaneous Raman s p e c t r o s c o p y can a r i s e .
(For more e x p e r i m e n t a l d e t a i l s see r e f . 3 ) .
Preparation PBS were i s o l a t e d a c c o r d i n g to the method p u b l i s h e d by
Nies
and Wehrmeyer ( 6 ) , except t h a t the pH of the phosphate b u f f e r was
6 and the p u r i f i c a t i o n on the s u c r o s e g r a d i e n t was
t w i c e . APC
done
was prepared v e r y s i m i l a r to a procedure d e s c r i b e d
by Füglistaller e t . a l . ( 7 ) . The a g g r e g a t i o n s t a t e of PC and APC mentation
by
sedi-
runs a c c o r d i n g to M a r t i n and Ames ( 8 ) . Both
was determined
bili-
p r o t e i n s were found to be t r i m e r i c . L i n k e r p e p t i d e s were not p r e s e n t as proved by a b s o r p t i o n s p e c t r a , s e d i m e n t a t i o n coe f f i c i e n t s and SDS-gel e l e c t r o p h o r e s i s .
Results As both the f i n g e r p r i n t
(1100 - 1300
bond s t r e t c h i n g r e g i o n (1500 - 1750
1
c u r ) and the double -1
cm )
have proved
to be
most s u i t a b l e f o r the i n v e s t i g a t i o n of chromophore geometry by CARS (3,4,5,9) or Resonance Raman S p e c t r o s c o p y both r e g i o n s a r e c o n s i d e r e d i n the f o l l o w i n g .
(10,11),
319
The
phycobilisome spectra
(PBS) a r e dominated by t h e c o n t r i b u
t i o n of t h e c o n s t i t u e n t phycocyanin t r i m e r s
(PC) and a r e t h e r e
f o r e r a t h e r s i m i l a r t o t h e s p e c t r a of t h e l a t t e r . Most of t h e v i b r a t i o n a l frequencies accuracy, 1200
a r e reproduced w i t h i n
experimental
though w i t h d i f f e r e n t i n t e n s i t i e s ( e s p e c i a l l y between 1
1
and 1300 cm- ) ( f i g 1 ) . At 1623 c n r an a d d i t i o n a l weak
peak i s found i n PBS, which i s due t o a l l o p h y c o c y a n i n
(APC).
A second i n t e r e s t i n g f e a t u r e i s t h a t f o r t h e pronounced bands a s m a l l e r w i d t h i s found i n PBS than i n PC. I t p o s s i b l y i n d i c a t e s a lower s t r u c t u r a l inhomogeneity i n PBS due t o h i g h e r a g g r e g a t i o n and t h e p r e s e n c e of l i n k e r
1200
1220
1210 12I1H Wavenumber s
1280
1500
1550
peptides.
1600 Vevcnumbers
IGS0
1/(HI
1 Λ.Π
F i g . 1: Resonance-enhanced CARS s p e c t r a of p h y c o b i l i s o m e s , phycocyanin t r i m e r s and a l l o p h y c o c y a n i n t r i m e r s from M a s t i g o c l a d u s l a m i n o s u s . Pump wavelength 640 nm.
320
APC s p e c t r a a r e v e r y d i f f e r e n t from PC s p e c t r a ( s e e a l s o 1
r e f . 1 0 ) . The 1257 cm" are absent
and 1287 cm"
1
bands observed 1
dominant peak i n PC i s found a t 1649 cm" 1
«1631 and »1662 cm"
1
«1635 cm"
with shoulders at
. I n APC s p e c t r a , however, the dominant
peak i s found a t 1625 cm" 1
i n PC
i n APC. I n the double bond s t r e t c h i n g r e g i o n the
with a strong shoulder a t 1
and a weak one a t «1650
cm" .
Discussion According
to p r e l i m i n a r y normal mode c a l c u l a t i o n s u s i n g the
coordinates provided
by X-ray a n a l y s i s
found between 1200 and 1300 cm
-1
( 1 2 , 1 3 ) , the bands
r e p r e s e n t e x t e n s i v e l y mixed
C-C s t r e t c h / C-H i n plane bending modes and can p r e s e n t l y not be i n t e r p r e t e d i n a s t r a i g h t - f o r w a r d manner. The band 1
around 1585 cm"
i s r e l a t e d to C=C s t r e t c h i n g v i b r a t i o n s i n
the r i n g B/C fragment. That change i n frequency,
t h i s band e x h i b i t s n e a r l y no
most l i k e l y i n d i c a t e s a c o n s e r v a t i o n of
the geometry of t h i s p a r t of the chromophore. The C=C double bonds of the methine b r i d g e s between r i n g and r i n g C-D,
A-B
on the o t h e r hand, s h o u l d g i v e r i s e t o l o c a l i z e d 1
modes w i t h f r e q u e n c i e s between 1650 and 1700 cm" .
The C=0
s t r e t c h mixes w i t h the C=C s t r e t c h i n r i n g D to produce v i b r a 1
t i o n a l f r e q u e n c i e s around 1630 cm" .
The a c t u a l v a l u e of the
c a l c u l a t e d f r e q u e n c i e s i s found to be s e n s i t i v e to s t r u c t u r a l changes. From t h e s e t h e o r e t i c a l r e s u l t s and the comparison of CARS s p e c t r a of model compounds (9) w i t h the p r e s e n t e d
CARS s p e c t r a
a s i g n i f i c a n t d i f f e r e n c e i n chromophore geometry and/or Π-electron
d i s t r i b u t i o n between PC and APC must be p o s t u l a t e d .
I n t e r p r e t a t i o n of e.g. f l u o r e s c e n c e decay d a t a of APC
should
t h e r e f o r e not be based on s t r u c t u r a l i n f o r m a t i o n g a i n e d PC.
from
321
A d d i t i o n a l measurements on antenna pigments and a more e l a b o r a t e normal mode a n a l y s i s are i n p r e p a r a t i o n .
Acknowledgement F i n a n c i a l support by Deutsche (SFB 143)
Forschungsgemeinschaft
and Fonds der Chemie i s g r a t e f u l l y acknowledged.
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