Effects of phospholipase A2 digestion on the carotenoid and bacteriochlorophyll components of the light-harvesting complexes in Rhodobacter sphaeroides chromatophores.
The instantaneous electrochromic response of carotenoids associated with the B800-850 light-harvesting complex of Rhodobacter sphaeroides has been used widely as an intrinsic probe of membrane potential. In the present study, the structural basis for this phenomenon was examined by phospholipase A2 digestion of chromatophores from R. sphaeroides strain NF57G, containing B800-850 as the sole pigment-protein complex. The major phospholipase-induced alterations of the overall carotenoid absorption spectrum were characterized by an absorbance loss and a blue shift that were accompanied by a decrease in absorbance at 800 nm and a red shift in the B850 absorbance band. In wild-type chromatophores, the electrochromic carotenoid response induced by both flash illumination and a K+ diffusion potential was diminished by approximately 60% after 1 h of digestion. The initial loss of the carotenoid response was correlated specifically to the hydrolysis of phosphatidylethanolamine, and was shown to arise from effects exerted directly upon the electrochromically active carotenoid pool, possibly by alterations in the spatial relationship between the field-sensitive carotenoids and the polarizing permanent field. In phospholipase A2-digested NF57G preparations in which the B800 band was diminished by nearly half and the carotenoid response was abolished, no significant changes in the efficiency of energy transfer from carotenoids to bacteriochlorophyll were detected at 77 K, suggesting that the electrochromically active carotenoids are not energetically linked to B800 bacteriochlorophyll.
Olivera
L M
LM
Department of Molecular Biology and Biochemistry, Rutgers University, Piscataway, New Jersey 08855-1059.
Niederman
R A
RA
D001429:Bacteriochlorophylls / Q000378:metabolism*; D002338:Carotenoids / Q000378:metabolism*; D004579:Electron Transport; D007425:Intracellular Membranes / Q000187:drug effects / Q000378:metabolism* / Q000528:radiation effects; D045342:Light-Harvesting Protein Complexes; D008055:Lipids / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy; D010741:Phospholipases A / Q000494:pharmacology*; D054467:Phospholipases A2; D010743:Phospholipids / Q000032:analysis; D045322:Photosynthetic Reaction Center Complex Proteins / Q000187:drug effects / Q000378:metabolism* / Q000528:radiation effects; D011188:Potassium / Q000494:pharmacology; D012242:Rhodobacter sphaeroides / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D013053:Spectrophotometry
D001429:Bacteriochlorophylls; D045342:Light-Harvesting Protein Complexes; D008055:Lipids; D010743:Phospholipids; D045322:Photosynthetic Reaction Center Complex Proteins; D002338:Carotenoids; D010741:Phospholipases A; D054467:Phospholipases A2; D011188:Potassium