Sauer, K.

Scheer, Hugo

Schneider, S.

English

Universität München: Elektronischen Publikationen

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 
DE G 
Walter de Gruyter · Berlin · New York 1988 
CONTENTS 
L i s t of P a r t i c i p a n t s X I I I 
SECTION I . ORGANIZATION: BIOCHEMICAL METHODS 
I n t r o d u c t i o n : 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. L i p s c h u l t z and F.X. Cunningham Jr 11 
On the S t r u c t u r e of Photosystem II-Phycobilisome Complexes 
of Cyanobacteria 
by E. Mörschel and G.-H. Schatz 21 
Stru c t u r e of Cryptophyte Photosynthetic Membranes 
by W. Wehrmeyer 35 
S t r u c t u r a l and Phylogenetic Relationships of Phycoerythrins 
from Cyanobacteria, Red Algae and Cryptophyceae 
by W. S i d l e r and H. Zuber 49 
I s o l a t i o n and C h a r a c t e r i z a t i o n of the Components of the 
Phycobilisome from Mastigocladus laminosus and Cross-
l 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 M o d i f i c a t i o n 
by R. Fischer, S. Siebzehnrlibl and H. Scheer 71 
Photochromic Prope r t i e s of C-Phycocyanin 
by G. Schmidt, S. Siebzehnrübl, R. Fischer and H. Scheer 77 
Concerning the Relat i o n s h i p of L i g h t Harvesting B i l i -
p r o t e i n s t o Phycochromes i n Cyanobacteria 
by W. Kufer 89 
Subunit S t r u c t u r e and Reassembly of the Light-Harvesting 
Complex from Rhodospiri Hum rubrum G9+ 
by R. Ghosh, ThT~Rosatzin ancTR. Baclofen 93 
Primary S t r u c t u r e Analyses of B a c t e r i a l Antenna Polypeptides 
- C o r r e l a t i o n of Aromatic Amino Acids w i t h Spectral 
Properties - S t r u c t u r a l S i m i l a r i t i e s w i t h Reaction Center 
Polypeptides 
by R.A. Brunisholz and H. Zuber 103 
VIII 
The S t r u c t u r e o f the "Core1* of the Purple B a c t e r i a l Photo-
s y n t h e t i c Unit 
by D.J. Dawkins, L.A. Ferguson and R.J. Cogdell 115 
A Comparison o f the Ba c t e r i o c h l o r o p h y l 1 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 B a c t e r i o c h l o r o p h y l 1 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 
Ligh t - H a r v e s t i n g 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 C h a r a c t e r i z a t i o n of a Chloro p h y l l a/c-Hetero-
xanthin/Diadinoxanthin Light-Harvesting Complex from 
P l e u r o c h l o r i s meiringensis (Xanthophyceae) 
by C. Wilhelm, C. Bliche I and B. Rousseau 167 
The Antenna Components of Photosystem I I w i t h 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 C r y s t a l S t r u c t u r e of C-Phycocyanin and 
Polarized O p t i c a l Spectra of Single C r y s t a l s 
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 I n v e s t i g a t i o n of Purple 
B a c t e r i a l B800-850 and RC-B875 Complexes 
by W. Welte, Τ. Wacker and A. Becker 201 
S t r u c t u r e of the Light-Harvesting Chlorophyll a/b-Protein 
Complex from Chloroplast Membranes 
by W. Kühlbrandt 211 
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 B a c t e r i a l Antenna Complexes 
by G. Drews 233 
IX 
The Use of Mutants t o Inv e s t i g a t e the Organization of the 
Photosynthetic Apparatus of Rhodobacter sphaeroides 
by C.N. Hunter and R. van Grondel le 7 7 . 7 7 247 
Mechanisms of P l a s t i d and Nuclear Gene Expression During 
Thylakoid Membrane Biogenesis i n 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 Or i e n t a t i o n of Pigments i n Phycobilisomes 
and t h e i r Subunits 
by L. Juszcak, N.E. Geacintov, B.A. Z i l i n s k a s 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. S z a l o n t a i , 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 
Op t i c a l Absorption and C i r c u l a r Dichroism of Bacteriochlorophyl1 
Oligomers i n T r i t o n 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 F i e l d 
on Antenna Complexes from Rps. acidophila 7050 - The 
Temperature Dependence of tfie CarotenoTiTTrTpTet State 
Prop e r t i e s 
by J. U l l r i c h , J.U. v. Schütz and H.C. Wolf 339 
E f f e c t o f Li t h i u m Dodecyl S u l f a t e on Β 800-850 Antenna 
Complexes from Rhodopseudomonas acidophila: A Resonance 
Raman Study 
by B. Robert and H. Frank 349 
χ 
B a c t e r i o c h l o r o p h y l 1 a/b i n Antenna Complexes of Purple Bacteria 
by Β. Robert, A. Vermeglio, R. Steiner, Η. Scheer and Μ. Lutz 355 
B a c t e r i o c h l o r o p h y l l c Aggregates i n Carbon T e t r a c h l o r i d e 
as Models f o r Chlo r o p h y l l Organization i n Green Photo-
s y n t h e t i c B acteria 
by J.M. Olson and J.P. Pedersen 365 
O r i e n t a t i o n o f t he 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 
E x c i t a t i o n Energy Transfer i n Photosynthesis 
by R. van Grondelle and V. Sundström 403 
Fluorescence Spectroscopy of Allophycocyanin Complexes from 
Synechococcus 6301 S t r a i n AN112 
by P.Maxson, KTTauer and A.N. Glazer 439 
Picosecond Energy Transfer K i n e t i c s i n Allophycocyanin 
Aggregates from Mastigocladus laminosus 
by E. Bittersmann, W. Reuter, W. Wehrmeyer and A.R. Holzwarth 451 
Picosecond Time-Resolved Energy Transfer K i n e t i c s 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-Phyco-
cyanin Trimers and the Constituent Subunits 
by S. Schneider, P. Gei s e l h a r t , F. Baumann, S. Siebzehnrübl, 
R. Fischer and Η. Scheer 469 
E f f e c t of P r o t e i n Environment and Exc i t o n i c Coupling on the 
Excited-State Pro p e r t i e s of the Bilinchromophores i n 
C-Phycocyanin 
by S. Schneider, Ch. Scharnagl, M. Dürring, T. Schirmer and 
W. Bode 483 
E x c i t a t i o n Energy M i g r a t i o n i n C-Phycocyanin Aggregates 
I s o l a t e d from Phormidium luridum: P r e d i c t i o n s from the 
Förster's I n d u c t i v e Resonance Theory 
by J. Grabowski and G.S. Björn 491 
XI 
Energy Tra n s f e r Calculations f o r two C-Phycocyanins Based on 
Refined X-Ray Crystal S t r u c t u r e Coordinates of Chromophores 
by K. Sauer and H. Scheer 507 
Energy Transfer i n Light-Harvesting Antenna o f Purple B a c t e r i a 
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 
E x c i t a t i o n Energy Transfer i n the Light-Harvesting Antenna o f 
Photosynthetic Purple B a c t e r i a : The Role of the Long-Wave-
Length Absorbing Pigment B896 
by R. van Grondelle, H. Bergström, V. Sundström, R.J. van Dorssen, 
M. Vos and C.N. Hunter 519 
The Function o f Chlorosomes i n Energy Transfer in Green Photo-
s y n t h e t i c Bacteria 
by R.J. van Dorssen, M. Vos and J. Amesz 531 
Energy Transfer i n Chloroflexus aurantiacus: E f f e c t s of 
Temperature and TSaerobic^onönHons 
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 o f Optical Spectra of Bacteriochlorophyl 1 
Antenna Complexes 
by R.M. P e a r l s t e i n 555 
Time Resolution and K i n e t i c s o f "F680" a t Low Temperatures i n 
Spinach Chloroplasts 
by R. Knox and S. L i n 567 
Picosecond Studies of Fluorescence and Absorbance Changes i n 
Photosystem I I P a r t i c l e s from Synechococcus Sp. 
by A.R. Holzwarth, G.H. Schatz and H. Brock 579 
Analysis of E x c i t a t i o n 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 
ENERGY TRANSFER CALCULATIONS FOR TWO C-PHYCOCYANINS BASED ON 
REFINED X-RAY CRYSTAL STRUCTURE COORDINATES OF CHROMOPHORES 
K. Sauer 
Laboratory of Chemical Biodynamics, University of C a l i f o r n i a , 
Berkeley, CA94720, USA 
H. Scheer 
Botanisches I n s t i t u t der Universität, Menzinger Str. 67, D-8000 
München 19, FRG 
Photosynthetic antennas are capable of e f f i c i e n t l i g h t absorp­
t i o n and e x c i t a t i o n energy transfer to the reaction centers 
over distances of several tens of nanometers. Light capture i s 
re a l i z e d by a v a r i e t y of strongly absorbing chromophores embed­
ded i n a pro t e i n matrix. These chromophores have absorption 
maxima that are t y p i c a l l y closely spaced. Energy tr a n s f e r pro­
ceeds i n a stepwise fashion among these chromophores. For these 
steps, two extreme mechanisms are generally considered (1). 
Weak exciton coupling i s operative over short distances ( t y p i c ­
a l l y £lnm), and Förster trans f e r remains e f f i c i e n t over much 
longer distances ( t y p i c a l l y ^6nm). In both cases, o r i e n t a t i o n 
and distances among the chromophores are c r i t i c a l parameters, 
on which l i t t l e information i s available i n most antenna sys­
tems . 
C-Phycocyanin (PC) i s one of the few cases where the necessary 
s t r u c t u r a l information i s known. Based on the preliminary 
c r y s t a l s t r u c t u r e of PC from the cyanobacterium Agmenellum 
quadruplicatum (2), rate constants f o r pairwise energy tr a n s f e r 
and the derived sets of d i f f e r e n t i a l equations have been deter­
mined (3) by using exclusively the Förster model (4). The f a i r 
agreement with experimentally determined absorption and 
Photosynthetic Light-Harvesting Systems 
© 1988 Walter de Gruyter & Co., Berlin · New York 
508 
Transfer Step New 
Coordinates 
Old 
ß-Subunit 
1B155 — > 1B84 34 4 (7 .6) 28 . 5 (6. 3) 
aß-Monomer 
1A84 --> 1B84 13 9 (10 .6) 0. 037 (0. 028) 
1B155 --> 1A84 2 15 (0 .52) 3 .44 (0. 84) 
(aß)3-Trimer 
3A84 --> 1B84 1527 (1160 . ) 59 .0 (44. 8) 
2B84 — > 1B84 8 8 (8 .8) 46 .2 (46. 2) 
2A84 --> 1B84 2 14 (1 .62) 0 .18 (0. 138) 
2B155 --> 1A84 2 4 (0 .58) 16 .9 (4. 1) 
2A84 --> 1A84 ι 25 (1 .25) 0 .42 (0. 42) 
3B155 --> 1B84 0 82 (0 .18) 6 . 2 (1. 37) 
2B155 --> 1B84 0 59 (0 . 131) 1 . 55 (0. 35) 
2B155 --> 1B155 0 29 (0 .29) 0 . 37 (0. 37) 
3B155 --> 1A84 0 092 (0 .023) 0 .054 (0. 0131) 
(aß)6-Hexamer 
4A84 — > 1A84 391 (391 . ) 185.4 (185. 4) 
6B155 — > 1B155 186 3 (186 .3) 174.8 (174. 8) 
6B155 --> 1A84 68 8 (16 .7) 20. 7 (5. 0) 
6A84 — > 1B84 40 7 (30 .9) 1. 64 (1 . 24) 
6B84 --> 1B84 10« 6 (10 .6) 10. 9 (10. 9) 
6B155 --> 1B84 9 2 (2 .04) 2 .01 (0. 45) 
5B84 --> 1B84 9 0 (9 .0) 53 .6 (53. 6) 
4B84 — > 1B84 4 5 (4 .5) 10 .2 (10. 2) 
5B155 --> 1B84 3 8 (0 .83) 1 .11 (0. 244) 
4A84 --> 1B84 1 31 (0 .99) 0 . 059 (0. 045) 6A84 — > 1A84 o 94 (0 .94) 0 . 58 (0. 58) 
4B155 — > 1B84 0 79 (0 .176) 1 . 31 (0. 29) 
5A84 --> 1B84 0 72 (0 . 54) 0 .044 (0. 033) 
4B155 --> 1A84 0 55 (0 .134) 3 . 5 (0. 85) 
5B155 --> 1A84 0 52 (0 .127) 0 .0033 (0. 0009) 
5A 84 --> 1A84 0 37 (0 .37) 0 . 134 (0. 134) 
5B155 — > 1B155 0 0112 (0 .0112) 0 .035 (0. 035) 
4B155 --> 1B155 0 0094 (0 .0094) 0 . 045 (0. 045) 
509 
fluorescence k i n e t i c data suggested that t h i s model might be 
s u f f i c i e n t to describe energy transfer i n PC. 
However, more recent experimental r e s u l t s (5-7) i n d i c a t e d a 
c o n f l i c t . Whereas theory predicted transfer between the two 
chromophores of lowest energy, e.g. oc84 and 684, as the f a s t e s t 
process, wavelength-resolved decay k i n e t i c s suggested t h a t 
t r a n s f e r from the high-energy 3155 chromophore to the low-
energy ones contributes strongly to the fastest resolved comp­
onent of appx. 20-40 ps. 
The recent refinement of the c r y s t a l structures of t r i m e r i c PC 
from Mastigocladus laminosus and hexameric PC from A. ^uadru-
plicatum (8), prompted us to reinvestigate t h i s problem. The 
r e f i n e d structures gave a much better d e f i n i t i o n of the chromo­
phores, i n p a r t i c u l a r of the previously only poorly resolved 
3-84 chromophore. This resulted i n minor changes i n the 
distances, but considerable changes i n the o r i e n t a t i o n para­
meters. We have now recalculated the energy t r a n s f e r and shown 
that the Förster mechanism may not be s u f f i c i e n t to describe 
the system, and that strong coupling has to be invoked at l e a s t 
f o r three symmetrically equivalent pairs of chromophores. 
The rate constants f o r pairwise energy transfer according to 
Förster theory are l i s t e d i n Table 1 ("new") for revised coord­
inates of PC from A. jguadruplicatum. The rate constants f o r 
Table 1: Rate Constants f o r Förster Ex c i t a t i o n Transfer i n PC 
Rate constants calculated using the Förster inductive resonance 
tr a n s f e r expression and spectroscopic overlap i n t e g r a l s , as 
described previously (3), but using a r e f r a c t i v e index value 
n=1.34 (10). Results which are compared are based on the " o l d " 
(2) and "new" (8) coordinates. Rate constants f o r backward 
tra n s f e r are indicated i n parenthesis. For increasing extent of 
aggregation, values from the top of the table are to be i n c l ­
uded i n a cumulative manner. A l l other tra n s f e r constants are 
r e l a t e d by symmetry (2,8) to the ones shown. 
510 
Μ. laminosus, are very s i m i l a r (not shown) , i n agreement w i t h 
the very s i m i l a r structures. The largest differences are calc­
ulated f o r rates i n v o l v i n g chromophore 3 1 5 5 , whose conformation 
seems to be somewhat affected by c r y s t a l packing ( 9 ) . As comp­
ared with the rate constants based on the previous s t r u c t u r a l 
data Table 1 , " o l d " ) , changes up to a f a c t o r of 40 are obs­
erved. Although t h i s increase was largest f o r some of the minor 
rate constants, some of the f a s t e s t and hence most important 
ones are affected as w e l l . I n aggregates,a r a t e of greater than 
l p s * 1 i s calculated f o r tran s f e r between adjacent a-84 and ß-84 
chromophores situated on d i f f e r e n t monomeric u n i t s . This rat e 
i s now by f a r the f a s t e s t , which i s even more pronounced 
Table 2: Excited State Exponential Decay Time Constants 
Calculated f o r Förster E x c i t a t i o n Transfer i n C-Phycocyanin 
i ! ß-subunit 
1 
Monomer Trimer Hexamer 
I T , 1 2 4 2 1 0 . 3 7 0 . 3 6 
k . 1 4 5 2 2 7 . 5 
1 1 5 0 0 
1 
1 5 0 0 1 5 0 0 1 5 0 0 
Calculated values l i s t e d are time constants i n ps f o r the f a s t 
(Γί > and intermediate { f 2 ) components of a three exponential 
analysis, with k F input as 0.67 ns~ 1 f o r the rate constant of 
r a d i a t i v e and non-radiative decay corresponding to ' t o . 
Table 3: Exciton Energies of Pairwise I n t e r a c t i o n Calculated 
for Chromophores of C-Phycocyanin (only values ^5cm_1 l i s t e d ) 
; 
I 1Chromophores i 
TV* , cm- 1 ] * 
! ß-Subunit ! '1B155/1B84 1 7 . 3 
' (aß) -Monomer 
ι 
J1A84/1B84 5.3 
! (aß)3-Trimer 
i 
J1A84/2B84 56 . 0 
' (αβ)β-Hexamer I1A84/4A84 32.0 
;1B155/6B155 23.8 
i J1A84/6B155 10.8 
I 
! 
J1A84/6B84 9.1 
• Exciton energies c a l -
' culated from the r e -
! l a t i o n V±=Dk/8R3 
I (11), with o r i e n t -
! a t i o n f a c t o r s , k, and 
' interchromophore d i s -
1 tances, R, taken from 
• Schirmer, et a l (8). 
! Dipole strength (D, 
I i n Debye 2) determined 
I from the spectrosco-
' pic deconvolution 
! ( 3 ) , wi t h d i e l e c t r i c 
J constant n=1.34 (10). 
511 
because the o r i g i n a l l y second fas t e s t rate w i t h i n the trimer 
(ß-84->ß-84) i s s t r o n g l y decreased. 
The occurence of these f a s t rates raised the p o s s i b i l i t y that 
other mechanisms of energy transfer must be considered. This i s 
supported by c a l c u l a t i o n s of exciton i n t e r a c t i o n s using p o i n t -
dipole approximation (table 3). Pairwise i n t e r a c t i o n s are i n 
p a r t i c u l a r s u b s t a n t i a l f o r α84/β84 on adjacent monomers, and 
fo r α84/α84 on d i f f e r e n t trimer disks i n the hexamer. Similar 
r e s u l t s were reported using point-monopole approximation (12). 
Acknowledgment 
This work was supported by the Deutsche Forschungsgemeinschaft 
(SFB 143, Forschergruppe "Pflanzliche Tetrapyrrole", and by the 
CIP botany computer f a c i l i t i e s , H.S.), by the Alexander-von-
Humboldt S t i f t u n g , Bonn (award to K.S.) and by the Director, 
O f f i c e of Energy Research, Office of Basic Energy Sciences, 
D i v i s i o n of B i o l o g i c a l Energy Conversion of the U.S. Department 
of Energy under Contract DE-AC03-76SFO0098 (K.S.). 
References 
1. Sauer, Κ. 1975. I n : Bioenergetics of Photosynthesis 
(Govindjee, ed.). Academic Press New York, p. 115. 
2. Schirmer, T., R. Huber, Μ. Schneider, W. Bode, Μ. M i l l e r , 
M.L. Hackert. 1986. J.Mol.Biol. 188, 651. 
3. Sauer, Κ., Η. Scheer, P. Sauer. 1987. Photochem.Photobiol. 
46, 427. 
4. Förster, Τ. 1967. I n : Comprehensive Biochemistry, Vol 22, 
(M. F l o r k i n and E.H. Stotz, eds.). Elsevier Amsterdam, p.61. 
5. Holzwarth,A.R., J.Wendler, G.W.Suter. 1987. Biophys.J. 51,1. 
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7. Schneider, S., P. Geiselhart, S. Siebzehnrübl, R. Fischer, 
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Energy transfer calculations for two C-phycocyanins based on refined x-ray crystal structure coordinates of chromophores

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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 DE G Walter de Gruyter · Berlin · New York 1988 CONTENTS L i s t of P a r t i c i p a n t s X I I I SECTION I . ORGANIZATION: BIOCHEMICAL METHODS I n t r o d u c t i o n : 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. L i p s c h u l t z and F.X. Cunningham Jr 11 On the S t r u c t u r e of Photosystem II-Phycobilisome Complexes of Cyanobacteria by E. Mörschel and G.-H. Schatz 21 Stru c t u r e of Cryptophyte Photosynthetic Membranes by W. Wehrmeyer 35 S t r u c t u r a l and Phylogenetic Relationships of Phycoerythrins from Cyanobacteria, Red Algae and Cryptophyceae by W. S i d l e r and H. Zuber 49 I s o l a t i o n and C h a r a c t e r i z a t i o n of the Components of the Phycobilisome from Mastigocladus laminosus and Cross-l 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 M o d i f i c a t i o n by R. Fischer, S. Siebzehnrlibl and H. Scheer 71 Photochromic Prope r t i e s of C-Phycocyanin by G. Schmidt, S. Siebzehnrübl, R. Fischer and H. Scheer 77 Concerning the Relat i o n s h i p of L i g h t Harvesting B i l i -p r o t e i n s t o Phycochromes i n Cyanobacteria by W. Kufer 89 Subunit S t r u c t u r e and Reassembly of the Light-Harvesting Complex from Rhodospiri Hum rubrum G9+ by R. Ghosh, ThT~Rosatzin ancTR. Baclofen 93 Primary S t r u c t u r e Analyses of B a c t e r i a l Antenna Polypeptides - C o r r e l a t i o n of Aromatic Amino Acids w i t h Spectral Properties - S t r u c t u r a l S i m i l a r i t i e s w i t h Reaction Center Polypeptides by R.A. Brunisholz and H. Zuber 103 VIII The S t r u c t u r e o f the "Core1* of the Purple B a c t e r i a l Photo-s y n t h e t i c Unit by D.J. Dawkins, L.A. Ferguson and R.J. Cogdell 115 A Comparison o f the Ba c t e r i o c h l o r o p h y l 1 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 B a c t e r i o c h l o r o p h y l 1 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 Ligh t - H a r v e s t i n g 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 C h a r a c t e r i z a t i o n of a Chloro p h y l l a/c-Hetero-xanthin/Diadinoxanthin Light-Harvesting Complex from P l e u r o c h l o r i s meiringensis (Xanthophyceae) by C. Wilhelm, C. Bliche I and B. Rousseau 167 The Antenna Components of Photosystem I I w i t h 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 C r y s t a l S t r u c t u r e of C-Phycocyanin and Polarized O p t i c a l Spectra of Single C r y s t a l s 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 I n v e s t i g a t i o n of Purple B a c t e r i a l B800-850 and RC-B875 Complexes by W. Welte, Τ. Wacker and A. Becker 201 S t r u c t u r e of the Light-Harvesting Chlorophyll a/b-Protein Complex from Chloroplast Membranes by W. Kühlbrandt 211 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 B a c t e r i a l Antenna Complexes by G. Drews 233 IX The Use of Mutants t o Inv e s t i g a t e the Organization of the Photosynthetic Apparatus of Rhodobacter sphaeroides by C.N. Hunter and R. van Grondel le 7 7 . 7 7 247 Mechanisms of P l a s t i d and Nuclear Gene Expression During Thylakoid Membrane Biogenesis i n 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 Or i e n t a t i o n of Pigments i n Phycobilisomes and t h e i r Subunits by L. Juszcak, N.E. Geacintov, B.A. Z i l i n s k a s 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. S z a l o n t a i , 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 Op t i c a l Absorption and C i r c u l a r Dichroism of Bacteriochlorophyl1 Oligomers i n T r i t o n 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 F i e l d on Antenna Complexes from Rps. acidophila 7050 - The Temperature Dependence of tfie CarotenoTiTTrTpTet State Prop e r t i e s by J. U l l r i c h , J.U. v. Schütz and H.C. Wolf 339 E f f e c t o f Li t h i u m Dodecyl S u l f a t e on Β 800-850 Antenna Complexes from Rhodopseudomonas acidophila: A Resonance Raman Study by B. Robert and H. Frank 349 χ B a c t e r i o c h l o r o p h y l 1 a/b i n Antenna Complexes of Purple Bacteria by Β. Robert, A. Vermeglio, R. Steiner, Η. Scheer and Μ. Lutz 355 B a c t e r i o c h l o r o p h y l l c Aggregates i n Carbon T e t r a c h l o r i d e as Models f o r Chlo r o p h y l l Organization i n Green Photo-s y n t h e t i c B acteria by J.M. Olson and J.P. Pedersen 365 O r i e n t a t i o n o f t he 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 E x c i t a t i o n Energy Transfer i n Photosynthesis by R. van Grondelle and V. Sundström 403 Fluorescence Spectroscopy of Allophycocyanin Complexes from Synechococcus 6301 S t r a i n AN112 by P.Maxson, KTTauer and A.N. Glazer 439 Picosecond Energy Transfer K i n e t i c s i n Allophycocyanin Aggregates from Mastigocladus laminosus by E. Bittersmann, W. Reuter, W. Wehrmeyer and A.R. Holzwarth 451 Picosecond Time-Resolved Energy Transfer K i n e t i c s 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-Phyco-cyanin Trimers and the Constituent Subunits by S. Schneider, P. Gei s e l h a r t , F. Baumann, S. Siebzehnrübl, R. Fischer and Η. Scheer 469 E f f e c t of P r o t e i n Environment and Exc i t o n i c Coupling on the Excited-State Pro p e r t i e s of the Bilinchromophores i n C-Phycocyanin by S. Schneider, Ch. Scharnagl, M. Dürring, T. Schirmer and W. Bode 483 E x c i t a t i o n Energy M i g r a t i o n i n C-Phycocyanin Aggregates I s o l a t e d from Phormidium luridum: P r e d i c t i o n s from the Förster's I n d u c t i v e Resonance Theory by J. Grabowski and G.S. Björn 491 XI Energy Tra n s f e r Calculations f o r two C-Phycocyanins Based on Refined X-Ray Crystal S t r u c t u r e Coordinates of Chromophores by K. Sauer and H. Scheer 507 Energy Transfer i n Light-Harvesting Antenna o f Purple B a c t e r i a 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 E x c i t a t i o n Energy Transfer i n the Light-Harvesting Antenna o f Photosynthetic Purple B a c t e r i a : The Role of the Long-Wave-Length Absorbing Pigment B896 by R. van Grondelle, H. Bergström, V. Sundström, R.J. van Dorssen, M. Vos and C.N. Hunter 519 The Function o f Chlorosomes i n Energy Transfer in Green Photo-s y n t h e t i c Bacteria by R.J. van Dorssen, M. Vos and J. Amesz 531 Energy Transfer i n Chloroflexus aurantiacus: E f f e c t s of Temperature and TSaerobic^onönHons 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 o f Optical Spectra of Bacteriochlorophyl 1 Antenna Complexes by R.M. P e a r l s t e i n 555 Time Resolution and K i n e t i c s o f "F680" a t Low Temperatures i n Spinach Chloroplasts by R. Knox and S. L i n 567 Picosecond Studies of Fluorescence and Absorbance Changes i n Photosystem I I P a r t i c l e s from Synechococcus Sp. by A.R. Holzwarth, G.H. Schatz and H. Brock 579 Analysis of E x c i t a t i o n 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 ENERGY TRANSFER CALCULATIONS FOR TWO C-PHYCOCYANINS BASED ON REFINED X-RAY CRYSTAL STRUCTURE COORDINATES OF CHROMOPHORES K. Sauer Laboratory of Chemical Biodynamics, University of C a l i f o r n i a , Berkeley, CA94720, USA H. Scheer Botanisches I n s t i t u t der Universität, Menzinger Str. 67, D-8000 München 19, FRG Photosynthetic antennas are capable of e f f i c i e n t l i g h t absorp­t i o n and e x c i t a t i o n energy transfer to the reaction centers over distances of several tens of nanometers. Light capture i s re a l i z e d by a v a r i e t y of strongly absorbing chromophores embed­ded i n a pro t e i n matrix. These chromophores have absorption maxima that are t y p i c a l l y closely spaced. Energy tr a n s f e r pro­ceeds i n a stepwise fashion among these chromophores. For these steps, two extreme mechanisms are generally considered (1). Weak exciton coupling i s operative over short distances ( t y p i c ­a l l y £lnm), and Förster trans f e r remains e f f i c i e n t over much longer distances ( t y p i c a l l y ^6nm). In both cases, o r i e n t a t i o n and distances among the chromophores are c r i t i c a l parameters, on which l i t t l e information i s available i n most antenna sys­tems . C-Phycocyanin (PC) i s one of the few cases where the necessary s t r u c t u r a l information i s known. Based on the preliminary c r y s t a l s t r u c t u r e of PC from the cyanobacterium Agmenellum quadruplicatum (2), rate constants f o r pairwise energy tr a n s f e r and the derived sets of d i f f e r e n t i a l equations have been deter­mined (3) by using exclusively the Förster model (4). The f a i r agreement with experimentally determined absorption and Photosynthetic Light-Harvesting Systems © 1988 Walter de Gruyter & Co., Berlin · New York 508 Transfer Step New Coordinates Old ß-Subunit 1B155 — > 1B84 34 4 (7 .6) 28 . 5 (6. 3) aß-Monomer 1A84 --> 1B84 13 9 (10 .6) 0. 037 (0. 028) 1B155 --> 1A84 2 15 (0 .52) 3 .44 (0. 84) (aß)3-Trimer 3A84 --> 1B84 1527 (1160 . ) 59 .0 (44. 8) 2B84 — > 1B84 8 8 (8 .8) 46 .2 (46. 2) 2A84 --> 1B84 2 14 (1 .62) 0 .18 (0. 138) 2B155 --> 1A84 2 4 (0 .58) 16 .9 (4. 1) 2A84 --> 1A84 ι 25 (1 .25) 0 .42 (0. 42) 3B155 --> 1B84 0 82 (0 .18) 6 . 2 (1. 37) 2B155 --> 1B84 0 59 (0 . 131) 1 . 55 (0. 35) 2B155 --> 1B155 0 29 (0 .29) 0 . 37 (0. 37) 3B155 --> 1A84 0 092 (0 .023) 0 .054 (0. 0131) (aß)6-Hexamer 4A84 — > 1A84 391 (391 . ) 185.4 (185. 4) 6B155 — > 1B155 186 3 (186 .3) 174.8 (174. 8) 6B155 --> 1A84 68 8 (16 .7) 20. 7 (5. 0) 6A84 — > 1B84 40 7 (30 .9) 1. 64 (1 . 24) 6B84 --> 1B84 10« 6 (10 .6) 10. 9 (10. 9) 6B155 --> 1B84 9 2 (2 .04) 2 .01 (0. 45) 5B84 --> 1B84 9 0 (9 .0) 53 .6 (53. 6) 4B84 — > 1B84 4 5 (4 .5) 10 .2 (10. 2) 5B155 --> 1B84 3 8 (0 .83) 1 .11 (0. 244) 4A84 --> 1B84 1 31 (0 .99) 0 . 059 (0. 045) 6A84 — > 1A84 o 94 (0 .94) 0 . 58 (0. 58) 4B155 — > 1B84 0 79 (0 .176) 1 . 31 (0. 29) 5A84 --> 1B84 0 72 (0 . 54) 0 .044 (0. 033) 4B155 --> 1A84 0 55 (0 .134) 3 . 5 (0. 85) 5B155 --> 1A84 0 52 (0 .127) 0 .0033 (0. 0009) 5A 84 --> 1A84 0 37 (0 .37) 0 . 134 (0. 134) 5B155 — > 1B155 0 0112 (0 .0112) 0 .035 (0. 035) 4B155 --> 1B155 0 0094 (0 .0094) 0 . 045 (0. 045) 509 fluorescence k i n e t i c data suggested that t h i s model might be s u f f i c i e n t to describe energy transfer i n PC. However, more recent experimental r e s u l t s (5-7) i n d i c a t e d a c o n f l i c t . Whereas theory predicted transfer between the two chromophores of lowest energy, e.g. oc84 and 684, as the f a s t e s t process, wavelength-resolved decay k i n e t i c s suggested t h a t t r a n s f e r from the high-energy 3155 chromophore to the low-energy ones contributes strongly to the fastest resolved comp­onent of appx. 20-40 ps. The recent refinement of the c r y s t a l structures of t r i m e r i c PC from Mastigocladus laminosus and hexameric PC from A. ^uadru-plicatum (8), prompted us to reinvestigate t h i s problem. The r e f i n e d structures gave a much better d e f i n i t i o n of the chromo­phores, i n p a r t i c u l a r of the previously only poorly resolved 3-84 chromophore. This resulted i n minor changes i n the distances, but considerable changes i n the o r i e n t a t i o n para­meters. We have now recalculated the energy t r a n s f e r and shown that the Förster mechanism may not be s u f f i c i e n t to describe the system, and that strong coupling has to be invoked at l e a s t f o r three symmetrically equivalent pairs of chromophores. The rate constants f o r pairwise energy transfer according to Förster theory are l i s t e d i n Table 1 ("new") for revised coord­inates of PC from A. jguadruplicatum. The rate constants f o r Table 1: Rate Constants f o r Förster Ex c i t a t i o n Transfer i n PC Rate constants calculated using the Förster inductive resonance tr a n s f e r expression and spectroscopic overlap i n t e g r a l s , as described previously (3), but using a r e f r a c t i v e index value n=1.34 (10). Results which are compared are based on the " o l d " (2) and "new" (8) coordinates. Rate constants f o r backward tra n s f e r are indicated i n parenthesis. For increasing extent of aggregation, values from the top of the table are to be i n c l ­uded i n a cumulative manner. A l l other tra n s f e r constants are r e l a t e d by symmetry (2,8) to the ones shown. 510 Μ. laminosus, are very s i m i l a r (not shown) , i n agreement w i t h the very s i m i l a r structures. The largest differences are calc­ulated f o r rates i n v o l v i n g chromophore 3 1 5 5 , whose conformation seems to be somewhat affected by c r y s t a l packing ( 9 ) . As comp­ared with the rate constants based on the previous s t r u c t u r a l data Table 1 , " o l d " ) , changes up to a f a c t o r of 40 are obs­erved. Although t h i s increase was largest f o r some of the minor rate constants, some of the f a s t e s t and hence most important ones are affected as w e l l . I n aggregates,a r a t e of greater than l p s * 1 i s calculated f o r tran s f e r between adjacent a-84 and ß-84 chromophores situated on d i f f e r e n t monomeric u n i t s . This rat e i s now by f a r the f a s t e s t , which i s even more pronounced Table 2: Excited State Exponential Decay Time Constants Calculated f o r Förster E x c i t a t i o n Transfer i n C-Phycocyanin i ! ß-subunit 1 Monomer Trimer Hexamer I T , 1 2 4 2 1 0 . 3 7 0 . 3 6 k . 1 4 5 2 2 7 . 5 1 1 5 0 0 1 1 5 0 0 1 5 0 0 1 5 0 0 Calculated values l i s t e d are time constants i n ps f o r the f a s t (Γί > and intermediate { f 2 ) components of a three exponential analysis, with k F input as 0.67 ns~ 1 f o r the rate constant of r a d i a t i v e and non-radiative decay corresponding to ' t o . Table 3: Exciton Energies of Pairwise I n t e r a c t i o n Calculated for Chromophores of C-Phycocyanin (only values ^5cm_1 l i s t e d ) ; I 1Chromophores i TV* , cm- 1 ] * ! ß-Subunit ! '1B155/1B84 1 7 . 3 ' (aß) -Monomer ι J1A84/1B84 5.3 ! (aß)3-Trimer i J1A84/2B84 56 . 0 ' (αβ)β-Hexamer I1A84/4A84 32.0 ;1B155/6B155 23.8 i J1A84/6B155 10.8 I ! J1A84/6B84 9.1 • Exciton energies c a l -' culated from the r e -! l a t i o n V±=Dk/8R3 I (11), with o r i e n t -! a t i o n f a c t o r s , k, and ' interchromophore d i s -1 tances, R, taken from • Schirmer, et a l (8). ! Dipole strength (D, I i n Debye 2) determined I from the spectrosco-' pic deconvolution ! ( 3 ) , wi t h d i e l e c t r i c J constant n=1.34 (10). 511 because the o r i g i n a l l y second fas t e s t rate w i t h i n the trimer (ß-84->ß-84) i s s t r o n g l y decreased. The occurence of these f a s t rates raised the p o s s i b i l i t y that other mechanisms of energy transfer must be considered. This i s supported by c a l c u l a t i o n s of exciton i n t e r a c t i o n s using p o i n t -dipole approximation (table 3). Pairwise i n t e r a c t i o n s are i n p a r t i c u l a r s u b s t a n t i a l f o r α84/β84 on adjacent monomers, and fo r α84/α84 on d i f f e r e n t trimer disks i n the hexamer. Similar r e s u l t s were reported using point-monopole approximation (12). Acknowledgment This work was supported by the Deutsche Forschungsgemeinschaft (SFB 143, Forschergruppe "Pflanzliche Tetrapyrrole", and by the CIP botany computer f a c i l i t i e s , H.S.), by the Alexander-von-Humboldt S t i f t u n g , Bonn (award to K.S.) and by the Director, O f f i c e of Energy Research, Office of Basic Energy Sciences, D i v i s i o n of B i o l o g i c a l Energy Conversion of the U.S. Department of Energy under Contract DE-AC03-76SFO0098 (K.S.). References 1. Sauer, Κ. 1975. I n : Bioenergetics of Photosynthesis (Govindjee, ed.). Academic Press New York, p. 115. 2. Schirmer, T., R. Huber, Μ. Schneider, W. Bode, Μ. M i l l e r , M.L. Hackert. 1986. J.Mol.Biol. 188, 651. 3. Sauer, Κ., Η. Scheer, P. Sauer. 1987. Photochem.Photobiol. 46, 427. 4. Förster, Τ. 1967. I n : Comprehensive Biochemistry, Vol 22, (M. F l o r k i n and E.H. Stotz, eds.). Elsevier Amsterdam, p.61. 5. Holzwarth,A.R., J.Wendler, G.W.Suter. 1987. Biophys.J. 51,1. 6. Sandström, Α., Τ. G i l l b r o , V. Sundström, R. Fischer, Η. Scheer: Biochim.Biophys.Acta. ( i n press) 7. Schneider, S., P. Geiselhart, S. Siebzehnrübl, R. Fischer, and H. Scheer, c o n t r i b u t i o n to t h i s volume 8. Schirmer, Τ., W. Bode, R. Huber. 1987. J.Mol.Biol. 196, 677. 9. Dürring, Μ., R.Huber (1987) Private communication 10. Moog, R.S., A. Kuki, M.D. Fayer, S.G. Boxer. 1984. Biochemistry 23, 1564. 11. Tinoco, I , J r . 1963. Radiation Research 20, 133. 12. Schneider,S., C.Scharnagl, M.Dürring, Τ.Schirmer, W.Bode, c o n t r i b u t i o n to t h i s book. 

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In: Bioenergetics of Photosynthesis (Govindjee,

M.Dürring, Τ.Schirmer, W.Bode, contribution to this book.

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Energy transfer calculations for two C-phycocyanins based on refined x-ray crystal structure coordinates of chromophores

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