Role of the core region of the PufX protein in inhibition of reconstitution of the core light-harvesting complexes of Rhodobacter sphaeroides and Rhodobacter capsulatus.
PufX, the protein encoded by the pufX gene of Rhodobacter capsulatus and Rhodobacter sphaeroides, has been further characterized. The mature forms of these proteins contain 9 and 12 fewer amino acids, respectively, at the C-terminal end of the protein than are encoded by their pufX genes. To identify the portion of PufX responsible for inhibition of LH1 formation in reconstitution experiments, different regions (N-terminus and several core regions containing different lengths of the C-terminus) of Rb. sphaeroides and Rb. capsulatus PufX were chemically synthesized. Neither the N- nor C-terminal polypeptides of Rb. sphaeroides were inhibitory to LH1 reconstitution. However, all core segments were active, causing 50% inhibition at a concentration ratio of between 3:1 and 6:1 relative to the LH1 alpha-polypeptides whose concentrations were 3-4 microM. CD measurements indicated that the core segment containing 39 amino acids of Rb. sphaeroides PufX exhibited 47% alpha-helix in trifluoroethanol while the core segment containing 43 amino acids of Rb. capsulatus PufX exhibited 59 and 55% alpha-helix in trifluoroethanol and in 0.80% octylglucoside in water, respectively. Approximately 50% alpha-helix was also indicated by a PHD (Burkhard-Rost) structure prediction. Binding of bacteriochlorophyll to these PufX core segments is implicated.
Parkes-Loach
P S
PS
Department of Biochemistry, Molecular Biology, and Cell Biology, Hogan Hall, Room 2-100, Northwestern University, Evanston, Illinois 60208-3500, USA.
Law
C J
CJ
Recchia
P A
PA
Kehoe
J
J
Nehrlich
S
S
Chen
J
J
Loach
P A
PA
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000138:chemical synthesis / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D001429:Bacteriochlorophylls / Q000037:antagonists & inhibitors* / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D002942:Circular Dichroism; D045342:Light-Harvesting Protein Complexes; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000302:isolation & purification / Q000378:metabolism; D045322:Photosynthetic Reaction Center Complex Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D016354:Rhodobacter capsulatus / Q000737:chemistry / Q000378:metabolism*; D012242:Rhodobacter sphaeroides / Q000737:chemistry / Q000378:metabolism*; D013045:Species Specificity
D001426:Bacterial Proteins; D001429:Bacteriochlorophylls; D045342:Light-Harvesting Protein Complexes; D010446:Peptide Fragments; D045322:Photosynthetic Reaction Center Complex Proteins; C088749:PufX protein, Rhodobacter; C094744:light-harvesting complex 1, Rhodospirillum