Photosynthetica 2002, 40(4):517-522 | DOI: 10.1023/A:1024387632311

Substitutions of the Conserved Thr42 Increased the Roles of the ε-Subunit of Maize CF1 as CF1 Inhibitor and Proton Gate

Zhang-Lin Ni1, Da-Fu Wang1, Jia-Mian Wei1
1 Shanghai Institute of Plant Physiology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China

The conserved residue Thr42 of ε-subunit of the chloroplast ATP synthase of maize (Zea mays L.) was substituted with Cys, Arg, and Ile, respectively, through site-directed mutagenesis. The over-expressed and refolded ε-proteins were purified by chromatography on DEAE-cellulose and FPLC on mono-Q column, which were as biologically active (inhibiting Ca2+-ATPase activity and blocking proton gate) as the native ε subunit isolated from chloroplasts. The εT42C and εT42R showed higher inhibitory activities on the soluble CF1(-ε) Ca2+-ATPase than the εWT. The εT42I inhibited the Ca2+-ATPase activity of soluble CF1 and restored photophosphorylation activity of membrane-bound CF1 deficient in ε the most efficiently. Far-ultraviolet CD spectra showed that the portions of α-helix and β-sheet structures of the three mutants were somewhat different from εWT. Thus the conserved residue Thr42 may be important for maintaining the structure and function of the ε-subunit and the basic functions of the ε-subunit as far as an inhibitor of Ca2+-ATPase and the proton gate are related.

Additional key words: ATP synthase; circular dichroism; coupling factor 1; ε-subunit; photophosphorylation; Zea mays

Published: December 1, 2002  Show citation

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Ni, Z., Wang, D., & Wei, J. (2002). Substitutions of the Conserved Thr42 Increased the Roles of the ε-Subunit of Maize CF1 as CF1 Inhibitor and Proton Gate. Photosynthetica, 40(4), 517-522. doi: 10.1023/A:1024387632311
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