The light-harvesting polypeptides of Rhodopseudomonas sphaeroides R-26.1. I. Isolation, purification and sequence analyses.
Four low-molecular-mass polypeptides were isolated and purified from chromatophore membranes of Rhodopseudomonas sphaeroides blue-green mutant R-26.1 by a combination of gel filtration and ion-exchange chromatography in organic solvents. On dodecyl sulfate polyacrylamide gels, the purified polypeptides comigrate with bands LH-1, LH-2 and LH-3 known to be related to the antenna-pigment-protein complexes. The complete primary structures were elucidated by automated Edman degradation of the intact polypeptides and of overlapping C-terminal fragments obtained after chemical cleavage at tryptophan and methionine residues. The C-termini were verified by hydrazinolysis and, in one case where an overlapping C-terminal fragment could not be obtained, by digestion with carboxypeptidase A. The four polypeptides show a tripartite structure: i.e. a polar N-terminal region is separated from a polar C-terminal region by a segment of about 21 predominantly hydrophobic amino-acid residues. All hydrophobic segments contain a characteristic conservative histidine residue. The C-terminal region is reduced to only a few amino acids in the two polypeptides which together form band LH-3, i.e. LH-3A and LH-3B. Their extended N-terminal region is rich in charged residues and contains an additional conserved histidine residue close to the beginning of the hydrophobic segment. These properties place LH-3A and LH-3B into subgroup (beta-polypeptides: B 870-beta and B 850-beta, respectively). LH-1 and LH-2 appear to form another subgroup (alpha-polypeptides: B 870-alpha and B 850-alpha, respectively) as suggested during a search for conservative elements within their sequences (structural basis for classification). N-Terminal analyses carried out with intact antenna-pigment-protein complexes revealed the following: (i) LH-1 and LH-3 are associated with the B 870 complex in Rp. sphaeroides 24.1 (wild type), (ii) the same polypeptides are almost exclusively present in chromatophore membranes of Rp. sphaeroides R-26, a blue-green mutant which absorbs at 870 nm, (iii) LH-2 and LH-3B are the constituent polypeptides of the B 800-850 complex of Rp. sphaeroides 2.4.1 and of the spectrally altered B 850 complex isolated from the blue-green mutant R-26.1 which absorbs at 860 nm. This mutant contains LH-2 and LH-3B along with LH-1 and LH-3A and apparently is able to form both types of antenna complexes.(ABSTRACT TRUNCATED AT 400 WORDS)
D000595:Amino Acid Sequence; D020130:Bacterial Chromatophores / Q000032:analysis; D001426:Bacterial Proteins / Q000032:analysis / Q000302:isolation & purification*; D002268:Carboxypeptidases; D043422:Carboxypeptidases A; D002848:Chromatography, DEAE-Cellulose; D003488:Cyanogen Bromide; D004591:Electrophoresis, Polyacrylamide Gel; D006834:Hydrazines; D006868:Hydrolysis; D007463:Iodobenzoates; D045322:Photosynthetic Reaction Center Complex Proteins; D012242:Rhodobacter sphaeroides / Q000032:analysis / Q000378:metabolism*
D001426:Bacterial Proteins; D006834:Hydrazines; D007463:Iodobenzoates; D045322:Photosynthetic Reaction Center Complex Proteins; C008154:2-iodosobenzoic acid; D002268:Carboxypeptidases; D043422:Carboxypeptidases A; D003488:Cyanogen Bromide