The role of extra fragment at the C-terminal of cytochrome b (Residues 421-445) in the cytochrome bc1 complex from Rhodobacter sphaeroides.
Sequence alignment of cytochrome b of the cytochrome bc1 complex from various sources reveals that bacterial cytochrome b contain an extra fragment at the C terminus. To study the role of this fragment in bacterial cytochrome bc1 complex, Rhodobacter sphaeroides mutants expressing His-tagged cytochrome bc1 complexes with progressive deletion from this fragment (residues 421-445) were generated and characterized. The cytbDelta-(433-445) bc1 complex, in which 13 residues from the C-terminal end of this fragment are deleted, has electron transfer activity, subunit composition, and physical properties similar to those of the complement complex, indicating that this region of the extra fragment is not essential. In contrast, the electron transfer activity, binding of cytochrome b, ISP, and subunit IV to cytochrome c1, redox potentials of cytochromes b and c1 in the cytbDelta-(427-445), cytbDelta-(425-445), and cytbDelta-(421-445) mutant complexes, in which 19, 21, or all residues of this fragment are deleted, decrease progressively. EPR spectra of the [2Fe-2S] cluster and the cytochromes b in these three deletion mutant bc1 complexes are also altered; the extent of spectral alteration increases as this extra fragment is shortened. These results indicate that the first 12 residues (residues 421-432) from the N-terminal end of the C-terminal extra fragment of cytochrome b are essential for maintaining structural integrity of the bc1 complex.
Liu
Xiaoying
X
Department of Biochemistry & Molecular Biology, Oklahoma State University, Stillwater, Oklahoma 74078, USA.
Yu
Chang-An
CA
Yu
Linda
L
D000595:Amino Acid Sequence; D055592:Biophysical Phenomena; D001703:Biophysics; D015153:Blotting, Western; D002856:Chromatophores / Q000737:chemistry; D003165:Complement System Proteins; D045303:Cytochromes b / Q000737:chemistry*; D004305:Dose-Response Relationship, Drug; D004578:Electron Spin Resonance Spectroscopy; D014450:Electron Transport Complex III / Q000737:chemistry*; D004583:Electrons; D004926:Escherichia coli / Q000378:metabolism; D017353:Gene Deletion; D006801:Humans; D007506:Iron-Sulfur Proteins / Q000737:chemistry; D013058:Mass Spectrometry; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D010084:Oxidation-Reduction; D011199:Potentiometry; D011485:Protein Binding; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D012242:Rhodobacter sphaeroides / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid
D007506:Iron-Sulfur Proteins; D003165:Complement System Proteins; D045303:Cytochromes b; D014450:Electron Transport Complex III