Electron transfer and stability of the cytochrome b6f complex in a small domain deletion mutant of cytochrome f.
The lumen segment of cytochrome f consists of a small and a large domain. The role of the small domain in the biogenesis and stability of the cytochrome b(6)f complex and electron transfer through the cytochrome b(6)f complex was studied with a small domain deletion mutant in Chlamydomonas reinhardtii. The mutant is able to grow photoautotrophically but with a slower rate than the wild type strain. The heme group is covalently attached to the polypeptide, and the visible absorption spectrum of the mutant protein is identical to that of the native protein. The kinetics of electron transfer in the mutant were measured by flash kinetic spectroscopy. Our results show that the rate for the oxidation of cytochrome f was unchanged (t(12) = approximately 100 micros), but the half-time for the reduction of cytochrome f is increased (t(12) = 32 ms; for wild type, t(12) = 2.1 ms). Cytochrome b(6) reduction was slower than that of the wild type by a factor of approximately 2 (t(12) = 8.6 ms; for wild type, t(12) = 4.7 ms); the slow phase of the electrochromic band shift also displayed a slower kinetics (t(12) = 5.5 ms; for wild type, t(12) = 2.7 ms). The stability of the cytochrome b(6)f complex in the mutant was examined by following the kinetics of the degradation of the individual subunits after inhibiting protein synthesis in the chloroplast. The results indicate that the cytochrome b(6)f complex in the small domain deletion mutant is less stable than in the wild type. We conclude that the small domain is not essential for the biogenesis of cytochrome f and the cytochrome b(6)f complex. However, it does have a role in electron transfer through the cytochrome b(6)f complex and contributes to the stability of the complex.
Gong
X S
XS
Department of Plant and Microbial Biology, University of California, Berkeley, California 94720, USA. xsg20@nature.berkeley.edu
Chung
S
S
Fernández-Velasco
J G
JG
D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000502:physiology; D016825:Chlamydomonas reinhardtii / Q000254:growth & development / Q000378:metabolism*; D003573:Cytochrome b Group / Q000378:metabolism*; D045346:Cytochrome b6f Complex; D003580:Cytochromes / Q000737:chemistry* / Q000235:genetics / Q000502:physiology; D045348:Cytochromes f; D004579:Electron Transport; D007700:Kinetics; D010788:Photosynthesis; D017434:Protein Structure, Tertiary; D017384:Sequence Deletion
D001426:Bacterial Proteins; D003573:Cytochrome b Group; D003580:Cytochromes; D045346:Cytochrome b6f Complex; D045348:Cytochromes f