Isolation and characterization of a novel membrane-bound cytochrome c553 from the strictly anaerobic phototroph, Heliobacillus mobilis

Abstract

Heliobacillus mobilis is a strictly anaerobic Gram-positive bacterium which contains a primitive Photosystem I-type reaction center. The membrane-bound cytochrome c553 from the heliobacterium suggested to be the immediate electron donor to the photooxidized pigment (P 798+) has been isolated and characterized. The heme protein was visualized as a major component with an apparent molecular size of 17 kDa in TMBZ-staining analysis of the membrane preparation and showed characteristic α (552.5 nm), β (522 nm), and Soret absorption (416 nm) peaks of a typical reduced c-type cytochrome in the partially purified sample. The internal 43 amino acid sequence of the electron donor was obtained by chemical agent and protease treatments followed by N-terminal sequencing of the resulting fragments. The internal sequence carries lots of lysine residues and a Cys-X-X-Cys-His sequence motif which are the characteristics of typical c-type cytochromes. The analysis of the sequence by FAST or FASTA program, however, did not show any significant similarity to other known heme proteins.