[27] Coupling Factors from Higher Plants

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Abstract

This chapter discusses the coupling factors from higher plants. The coupling device of energy transduction is essentially a proton translocating ATPase that is composed of two sectors, a membrane-bound ATPase that can readily be released to a soluble form and a membrane sector that conducts protons across the membrane. Proton ATPases from various organisms resemble each other in many respects. It is assumed the mitochondrial coupling device from plants should be similar to those from yeast and animals. Chloroplast coupling factor 1 (CF1) is the ATPase enzyme from chloroplasts. Modified procedures and new developments concerning CF1 and the membrane sector of the chloroplast coupling device are described in this chapter. The latent ATPase activity of CF1 can be activated by heat treatment or by trypsin digestion or by prolonged incubation at high concentrations of dithiothreitol. An alternative procedure for trypsin activation is to include 25 μg of trypsin in the ATPase assay medium. This chapter also discusses preparation of [γ-32P]ATP. Assay of Coupling Activity of CF1 and its preparation is described in the chapter. Purification and properties of a chloroplast proteolipid are also elaborated.

Original languageEnglish
Pages (from-to)301-313
Number of pages13
JournalMethods in Enzymology
Volume69
Issue numberC
DOIs
StatePublished - 1 Jan 1980

Funding

FundersFunder number
United States-Israel Binational Science Foundation

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