The Rhodobacter sphaeroides cytochrome c2 signal peptide is not necessary for export and heme attachment.
Rhodobacter sphaeroides cytochrome c2 (cyt c2) is a member of the heme-containing cytochrome c protein family that is found in the periplasmic space of this gram-negative bacterium. This exported polypeptide is made as a higher-molecular-weight precursor with a typical procaryotic signal peptide. Therefore, cyt c2 maturation is normally expected to involve precursor translocation across the cytoplasmic membrane, cleavage of the signal peptide, and covalent heme attachment. Surprisingly, synthesis as a precursor polypeptide is not a prerequisite for cyt c2 maturation because deleting the entire signal peptide does not prevent export, heme attachment, or function. Although cytochrome levels were reduced about threefold in cells containing this mutant protein, steady-state cyt c2 levels were significantly higher than those of other exported bacterial polypeptides which contain analogous signal peptide deletions. Thus, this mutant protein has the unique ability to be translocated across the cytoplasmic membrane in the absence of a signal peptide. The covalent association of heme with this mutant protein also suggests that the signal peptide is not required for ligand attachment to the polypeptide chain. These results have uncovered some novel aspects of bacterial c-type cytochrome biosynthesis.
Brandner
J P
JP
Department of Bacteriology, University of Wisconsin, Madison 53706.
Donohue
T J
TJ
D000595:Amino Acid Sequence; D001483:Base Sequence; D001692:Biological Transport; D003001:Cloning, Molecular; D003574:Cytochrome c Group / Q000737:chemistry / Q000378:metabolism*; D045362:Cytochromes c2; D006418:Heme / Q000378:metabolism; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D021382:Protein Sorting Signals / Q000378:metabolism*; D015183:Restriction Mapping; D012242:Rhodobacter sphaeroides / Q000737:chemistry*; D013329:Structure-Activity Relationship
D003574:Cytochrome c Group; D009838:Oligodeoxyribonucleotides; D021382:Protein Sorting Signals; D006418:Heme; D045362:Cytochromes c2