title: Cytochrome f from the Antarctic psychrophile, Chlamydomonas raudensis UWO 241: structure, sequence, and complementation in the mesophile, Chlamydomonas reinhardtii.
abstract: Although cytochrome f from the Antarctic psychrophile, Chlamydomonas raudensis UWO 241, exhibits a lower apparent molecular mass (34 kD) than that of the mesophile C. reinhardtii (41 kD) based on SDS-PAGE, both proteins are comparable in calculated molecular mass and show 79% identity in amino acid sequence. The difference in apparent molecular mass was maintained after expression of petA from both Chlamydomonas species in either E. coli or a C. reinhardtii DeltapetA mutant and after substitution of a unique third cysteine-292 to phenylalanine in the psychrophilic cytochrome f. Moreover, the heme of the psychrophilic form of cytochrome f was less stable upon heating than that of the mesophile. In contrast to C. raudensis, a C. reinhardtii DeltapetA mutant transformed with petA from C. raudensis exhibited the ability to undergo state transitions and a capacity for intersystem electron transport comparable to that of C. reinhardtii wild type. However, the C. reinhardtii petA transformants accumulated lower levels of cytochrome b ( 6 ) /f complexes and exhibited lower light saturated rates of O(2) evolution than C. reinhardtii wild type. We show that the presence of an altered form of cytochrome f in C. raudensis does not account for its inability to undergo state transitions or its impaired capacity for intersystem electron transport as previously suggested. A combined survey of the apparent molecular mass, thermal stability and amino acid sequences of cytochrome f from a broad range of mesophilic species shows unequivocally that the observed differences in cytochrome f structure are not related to psychrophilly. Thus, caution must be exercised in relating differences in amino acid sequence and thermal stability to adaptation to cold environments.
authors.affiliation: Department of Biology and The Biotron, University of Western Ontario, London, Canada, N6A 5B7.
authors.forename: Loreta
authors.initials: L
authors.lastname: Gudynaite-Savitch
authors.forename: Michael
authors.initials: M
authors.lastname: Gretes
authors.forename: Rachael M
authors.initials: RM
authors.lastname: Morgan-Kiss
authors.forename: Leonid V
authors.initials: LV
authors.lastname: Savitch
authors.forename: John
authors.initials: J
authors.lastname: Simmonds
authors.forename: Susanne E
authors.initials: SE
authors.lastname: Kohalmi
authors.forename: Norman P A
authors.initials: NP
authors.lastname: Hüner
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000864:Antarctic Regions; D029681:Arabidopsis Proteins / Q000235:genetics; D002696:Chlamydomonas / Q000235:genetics*; D016825:Chlamydomonas reinhardtii / Q000235:genetics; D002736:Chloroplasts / Q000235:genetics; D003001:Cloning, Molecular; D045348:Cytochromes f / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D004579:Electron Transport; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics; D005816:Genetic Complementation Test; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009154:Mutation; D011487:Protein Conformation; D017421:Sequence Analysis
chemical_list: D029681:Arabidopsis Proteins; C472828:PETA protein, Arabidopsis; D045348:Cytochromes f