FEBS Letters | |
Fragmentation of chloroplast coupling factor in dependence of bound nucleotides Preparation of a reconstitutionally active form of subunit δ | |
Engelbrecht, S.1  Junge, W.1  | |
[1] Abt. Biophysik, Fachbereich Biologie/Chemie, Universität Osnabrück, Barbarastr. 11, D-4500 Osnabrück, FRG | |
关键词: CF1; Nucleotide content; Phosphorylation; Photosynthesis; H +-ATPase; CF0CF1; chloroplast ATP synthase; CF1; chloroplast ATPase; CF0; chloroplast ATP synthase; proton-conducting part; CF1(-δ); CF1 lacking the δ-subunit; CF1(-ϵ); CF1 lacking the ϵ-subunit; CF1(-δϵ); CF1 lacking the δ- and ϵ- subunits; Mega 9; N-(D-gluco-2; 3; 4; 5; 6-pentahydroxylhexyl)-N-methylnonanamide; | |
DOI : 10.1016/0014-5793(87)80244-2 | |
学科分类:生物化学/生物物理 | |
来源: John Wiley & Sons Ltd. | |
【 摘 要 】
Previous studies on the ability of CF1, fragments to reconstitute photophosphorylation in CF1,-depleted thylakoids have shown that the degree of reconstitution was correlated with the presence of subunit δ in the fragment. This was taken as evidence that subunit δ was necessary for plugging the active proton channel CF0 [(1986) Eur. J. Biochem. 160, 635–643]. We questioned whether or not δ alone had this ability. In order to obtain δ we investigated the role of bound nucleotides in the stability of CF1. Starting from ammonium sulfate-precipitated CF1, we found that a low content of bound ADP (1 mol ADP/mol CF1) seemed to stabilize the β—δ interaction, while loosening the interaction between α,β and γ. By elution from an anion-exchange column in the presence of the nonionic surfactant Mega 9 we obtained β3δ and CF1(—δ) (both containing one ADP) or, after washing with alcohol/glycerol mixtures, β (nucleotide-free) and CF1/CF1(—ϵ). On the other hand, with a further 2 ADP and 2 ATP bound to CF1, (after incubation with excess ATP) the α-β-γ interaction was stabilized in such a way that subunit δ alone could be isolated from the complex. Subunit δ, when isolated by this procedure and added back to CF1-depleted thylakoids, reconstituted a high rate of photophosphorylation.
【 授权许可】
Unknown
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