Structure-based sequence alignment of several

Table S1. Kinetic parameters of NADPH diaphorase reaction for several FNRs and the dissociation constants for complexes with NADP+ FNR X. axonopodisa peaa E.colia R. capsulatus P. putidad a

Km ( M) 10.8±0.5 15.3±4.3 8.3±1.3 80b 4.79±0.08 3.97±0.04

kcat (s-1) 121.9±1.8 374.3±18 38.2±3.5 7.2 b 4.77±0.17 4.510.06

kcat/ Km ( M-1 s-1) 11.3 24.5 4.6 0.09 1.03±0.02 1.14±0.08

Kd ( M) 9.7±0.4 10.9±3.1 5.9±0.6 222±5c -

Data from Tondo M.L. et al., PLoS One 2011; 6: e27124. Data from Bittel C. et al., FEBS Lett 2003; 553: 408–412. Data from Bortolotti A. et al., BBA 2009; 1794: 199-210. d Data from Yeom J. et al., J. Biochem 2009; 145: 481-491. Data for FprA (first line), homologue with the A. vinelandii FPR and FprB (second line), homologue with the E.coli FPR. b c

Figure S1 1

XacFP PaFPR AvFPR RcFPR StFPR EcFPR

(4b4d) (2qdx) (1a8p) (2bgi) (3fpk) (1fdr)

2

1

6

8

7

5

6

7

9

10

61 60 60 73 59 59

3

121 120 120 133 119 119

9

PWLSIIKDPETYERFDKVILTQGVRFVQDL-AYRDYFERELPQHEFLGDLLREKLLYYPA PFLSVIQDPETYERYEKVILVHGVRWVSEL-AYADFITKVLPEHEYFGDQVKEKLIYYPL PFMSLIQDPEVYERFEKVVLIHGVRQVNEL-AYQQFITEHLPQSEYFGEAVKEKLIYYPT PFASLMREPEAYEKFDEVIMMHACRTVAELeYGRQLVE-ALQEDPLIGELVEGKLKYYPT PYLSILQYGQDVARFKNLVLVHAARFAADL-SYLPLML-ELQQR------YEGKLRIQTV PYLSILRLGKDLDRFKNLVLVHAARYAADL-SYLPLMQ-ELEKR------YEGKLRIQTV 8

XacFPR(4b4d) PaFPR (2qdx) AvFPR (1a8p) RcFPR (2bgi) StFPR (3fpk) EcFPR (1fdr)

2

EEHLEFFSIKVPDGPLTSRLQHIQPGDKVLVGKKPTGTLLISDLHPGRNLYLLGTGTGLA EEHLEFFSIKVPDGPLTSRLQHLKEGDELMVSRKPTGTLVHDDLLPGKHLYLLSTGTGMA EEHLEFFSIKVQNGPLTSRLQHLKEGDELMVSRKPTGTLVTSDLLPGKHLYMLSTGTGLA DEELEFYSIKVPDGPLTSRLQHIKVGEQIILRPKPVGTLVIDALLPGKRLWFLATGTGIA NPNLEFYLVTVPQGKLSPRLAALKPGDEVQVVSDASGFFVLDEVPDCETLWMLATGTAIG NPDLEFYLVTVPDGKLSPRLAALKPGDEVQVVSEAAGFFVLDEVPHCETLWMLATGTAIG

4 XacFPR(4b4d) PaFPR (2qdx) AvFPR (1a8p) RcFPR (2bgi) StFPR (3fpk) EcFPR (1fdr)

4

-AFGAETVLEVRHWTDAYFSFTTTRDAGFRFENGQFVMIGLETET-RPLLRAYSIASANW sNLYTERVLSVHHWNDTLFSFKTTRNPGLRFKTGQFVMIGLEVDG-RPLMRAYSIASPNY sNLNVERVLSVHHWNDTLFSFKTTRNPSLRFENGQFVMIGLEVDG-RPLMRAYSIASPNY --PDAQTVTSVRHWTDTLFSFRVTRPQTLRFRSGEFVMIGLLDDNgKPIMRAYSIASPAW aDWVTGKVTKVQNWTDALFSLTVHAP-iNPFTAGQFTKLGLEIDG-ERVQRAYSYVNAPD aDWVTGKVTKVQNWTDALFSLTVHAP-vLPFTAGQFTKLGLEI----RVQRAYSYVNSPD 5

XacFPR(4b4d) PaFPR (2qdx) AvFPR (1a8p) RcFPR (2bgi) StFPR (3fpk) EcFPR (1fdr)

3

180 179 179 192 171 169

10

VTRETF--ANQGRLTELMADGRMQQTLGLPTLDPANDRFMICGSPQMLADLRSLLDS-RG VTREPF--RNQGRQTDLMRSGKLFEDIGLPPMNPQDDRAMICGSPSMLEETSAVLDS-FG VTRESF--HNQGRLTDLMRSGKLFEDIGLPPINPQDDRAMICGSPSMLDESCEVLDG-FG TTREEF--HHMGRITDNLASGKVFEDLGIAPMNPETDRAMVCGSLAFNVDVMKVLES-YG VSRENVpgSLTGRVPALIENGELEKAVGLP-MDKETSHVMLCGNPQMVRDTQQLLKEtRQ VSRETAagSLTGRIPALIESGELESTIGLP-MNKETSHVMLCGNPQMVRDTQQLLKEtRQ

237 236 236 249 230 230

11

XacFPR(4b4d) PaFPR (2qdx) AvFPR (1a8p) RcFPR (2bgi) StFPR (3fpk) EcFPR (1fdr)

FQTSPRIGTPGHYVFERAFVEK-LKISPRMGEPGDYLIERAFVEK-LKISPRMGEPGDYLIERAFVEK-LREG-ANSEPREFVVEKAFVGEgi MTKHLR-rRPGHMTAEHYW----MTKHLR-rRPGHMTAEHYW-----

259 258 258 272 248 248

Figure S1: Structure-based sequence alignment of several bacterial FPRs by DALI Server. Secondary structure for XacFPR stated by the SMM server. Pa, P. aeruginosa (subclass IA); Av, A. vinelandii (subclass IA); Rc, R. capsulatus (subclass IB); St, Salmonella typhimurium (subclass II); Ec, E. coli (subclass II). Residues of XacFPR that interact with FAD are pointed with an asterisk on top.