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Plant and Cell Physiology, 1985, Vol. 26, No. 3 505-514
© 1985

Article

Effect of Methanol on the Nucleotide Binding to High-Affinity Sites on Chloroplast Coupling Factor 1

Toru Hisabori and Hidehiro Sakurai

Department of Biology, School of Education, Waseda University Nishiwaseda, Shinjuku-ku, Tokyo 160, Japan

The effects of methanol on the nucleotide binding to isolated chloroplast coupling factor 1 (CF1) were investigated. Isolated CF1 has four kinds of nucleotide binding sites; a barely dissociable ADP-binding site (site A), two slowly exchangeable high-affinity sites with different affinities for ADP (sites B and C) which are not catalytic sites, and several low-affinity sites (Hisabori and Sakurai 1984). Methanol at 20% (v/v) slightly accelerated the binding of ADP to CF1 but did not influence the number of binding sites. Methanol at 10–24% (v/v) affected neither the total amounts of bound adenine nucleotides (~2.5 mol/mol CF1) nor the incorporation of labeled ADP from the medium (~1.5 mol/mol CF1 into the slowly exchangeable sites (sites A, B, C). These results indicate that no appreciable exchange of ADP occurred at site A at 10–24% (v/v) methanol and exclude the possibility of direct participation of nucleotide binding at this site in the regulation of ATPase. In 32% methanol, the amount of the labeled ADP bound increased, suggesting some exchange at site A. Methanol at 20% (v/v) greatly increased the affinities of sites B and C for ADP, CDP, GDP, UDP and PPi.

Conformational change of CF1 induced by the binding of nucleotides to site(s) B (and C) increased the resistance of CF1 to inactivation by methanol at high concentrations or by cold treatment.

(Received August 16, 1984; Accepted January 23, 1985)
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