Reconstitution of CF1-depleted thylakoid membranes with complete and fragmented chloroplast ATPase. The role of the delta subunit for proton conduction through CF0

Siegfried Engelbrecht, H Lill, Wolfgang Junge

Research output: Contribution to JournalArticleAcademicpeer-review

Abstract

Chloroplast ATPase (CF1) was isolated from spinach, pea and maize thylakoids by EDTA extraction followed by anion-exchange chromatography. CF1 was purified and resolved by HPLC into integral CF1, and CF1 lacking the delta & epsilon subunits: CF1(-delta) and CF1(-epsilon). Washing Mono-Q-bound CF1 with alcohol-containing buffers followed by elution without alcohol produced the beta subunit and in separate peaks CF1(-delta) and CF1(-epsilon). Elution from Mono Q in the presence of tenside yielded a beta delta fragment, CF1(-delta) and CF1(-delta epsilon). Chloroplasts were CF1-depleted by EDTA extraction. Reconstitution of photophosphorylation in these 'EDTA vesicles' was obtained by addition of CF1 and its fragments. CF1, CF1(-delta) and CF1(-delta epsilon) were active with cross-reactivity between spinach, pea and maize. delta-containing CF1 always reconstituted higher activities than delta-deficient CF1. The beta delta fragment and dicyclohexylcarbodiimide (DCCD)-inhibited CF1 also were reconstitutively active while beta and DCCD-inhibited CF1(-delta) were not. These results support the notion that subunit delta can function as a stopcock to the CF0 proton channel as proposed by Junge, W., Hong, Y. Q., Qian, L. P. and Viale, A. [(1984) Proc. Natl Acad. Sci. USA 81, 3078-3082].

Original languageEnglish
Pages (from-to)635-43
Number of pages9
JournalEuropean Journal of Biochemistry
Volume160
Issue number3
DOIs
Publication statusPublished - 3 Nov 1986

Keywords

  • Chloroplasts
  • Immunoelectrophoresis
  • Kinetics
  • Macromolecular Substances
  • Peptide Fragments
  • Plants
  • Proton-Translocating ATPases
  • Protons
  • Species Specificity
  • Journal Article
  • Research Support, Non-U.S. Gov't

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