A negatively charged N terminus in the alpha polypeptide inhibits formation of light-harvesting complex I in Rhodobacter capsulatus.
Light-harvesting complex I (LHI) of Rhodobacter capsulatus contains bacteriochlorophyll and carotenoids which are noncovalently bound to two different apoproteins (alpha and beta polypeptides) carrying oppositely charged N-terminal ends. The contribution of these charged segments to the assembly of LHI was studied with mutants having oppositely charged amino acids in the alpha or beta polypeptide. The influence of these mutations on the insertion and assembly process of the LHI complex was investigated by means of spectroscopic analysis of isolated intracytoplasmic membranes and pulse-chase experiments. Exchange of four positively charged amino acids to negatively charged amino acids on the N-terminal domain of the alpha subunit inhibited completely the assembly of the LHI complex. Although this mutant has no antenna, the reaction center is active and the cells were able to grow anaerobically in the light. Conversely, mutation of the four negatively charged amino acids of the N-terminal segment of the beta polypeptide did not prevent the assembly of the LHI complex, although the stability of the complex and the size of the photosynthetic unit were affected. The presence of the mutated beta polypeptide was confirmed by protein sequencing.
Stiehle
H
H
Institut für Biologie II, Albert-Ludwigs-Universität, Freiburg, Federal Republic of Germany.
Cortez
N
N
Klug
G
G
Drews
G
G
D000595:Amino Acid Sequence; D007526:Isoelectric Point; D045342:Light-Harvesting Protein Complexes; D046911:Macromolecular Substances; D008565:Membrane Proteins / Q000737:chemistry; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010084:Oxidation-Reduction; D045322:Photosynthetic Reaction Center Complex Proteins / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure; D016354:Rhodobacter capsulatus / Q000032:analysis / Q000502:physiology*; D013057:Spectrum Analysis; D013329:Structure-Activity Relationship
D045342:Light-Harvesting Protein Complexes; D046911:Macromolecular Substances; D008565:Membrane Proteins; D045322:Photosynthetic Reaction Center Complex Proteins