TYROSINE-162 OF THE PHOTOSYNTHETIC REACTION-CENTER L-SUBUNIT PLAYS A CRITICAL ROLE IN THE CYTOCHROME-C(2) MEDIATED REREDUCTION OF THE PHOTOOXIDIZED BACTERIOCHLOROPHYLL DIMER IN RHODOBACTER-SPHAEROIDES .1. SITE-DIRECTED MUTAGENESIS AND INITIAL CHARACTERIZATION

被引:33
作者
FARCHAUS, JW [1 ]
WACHTVEITL, J [1 ]
MATHIS, P [1 ]
OESTERHELT, D [1 ]
机构
[1] MAX PLANCK INST BIOCHEM,DEPT MEMBRANE BIOCHEM,KLOPFERSPITZ 18A,D-82143 MARTINSRIED,GERMANY
关键词
D O I
10.1021/bi00091a044
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Five site-directed mutants were engineered to substitute phenylalanine, serine, leucine, methionine, and glycine for tyrosine residue 162 of the pufL gene in Rhodobacter (R.) sphaeroides. Each of the mutations and the wild-type (WT) genes was expressed in the R. sphaeroides puf deletion strain PUFDELTALMX21/3. Initial characterization revealed that all of the mutants were photoheterotrophically competent but that L162G and L162S were impaired. The amounts of mutant reaction centers expressed, the spectral characteristics, and the rates of intraprotein electron transfer and turnover were similar to the values obtained for WT. Kinetic measurements of photooxidized special pair rereduction mediated by the physiological donor cytochrome c2 in intact chemoheterotrophically grown cells revealed that the fast phase was abolished in all mutants and that the overall kinetics of rereduction was drastically slowed. It is concluded that L162Y plays a vital role in facilitating the rapid rereduction of the photooxidized bacteriochlorophyll dimer in R. sphaeroides.
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页码:10885 / 10893
页数:9
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