title: Properties of the cyanobacterial coupling factor ATPase from Spirulina platensis. I. Electrophoretic characterization and reconstitution of photophosphorylation.
abstract: The coupling factor ATPase (F1) from photosynthetic membranes of the cyanobacterium Spirulina platensis was purified to homogeneity by a combination of ion-exchange chromatography and sucrose density gradient centrifugation. The ATPase activity of purified Spirulina F1 is latent but can be elicited by trypsin treatment, resulting in specific activities (CaATPase) of 27-37 mumol Pi min-1 mg protein-1. On denaturing sodium dodecyl sulfate-polyacrylamide gradient gels, Spirulina F1 is resolved into five subunits with molecular weights of 53,400, 51,600, 36,000, 21,100, and 14,700, similar to the molecular weights of the subunits of spinach chloroplast coupling factor (CF1). As determined by native polyacrylamide gradient gel electrophoresis, the molecular weight of the Spirulina F1 holoenzyme was estimated to be 320,000, somewhat smaller than the estimated molecular weight of spinach CF1 (392,000). Spirulina F1 was shown to be an active coupling factor by its ability to reconstitute phenazine methosulfate-dependent cyclic photophosphorylation in membrane vesicles which had been depleted of coupling factor content by 2 M NaBr treatment. We estimate the Spirulina F1 content of membrane vesicles to be 1 F1 per 830 chlorophylls or 0.12 mol F1 mol P700(-1), based on the specific ATPase activities of the membrane vesicles and the purified Spirulina F1, the molecular weight of F1, and the P700 content of the vesicles.
authors.forename: D B
authors.initials: DB
authors.lastname: Hicks
authors.forename: C F
authors.initials: CF
authors.lastname: Yocum
mesh_terms: D002499:Centrifugation, Density Gradient; D000458:Cyanobacteria / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel; D008970:Molecular Weight; D010785:Photophosphorylation; D006180:Proton-Translocating ATPases / Q000302:isolation & purification*; D012997:Solvents
chemical_list: D012997:Solvents; D006180:Proton-Translocating ATPases