Isolation of Subunits of Coupling Factor 1 from Maize and Spinach Chloroplasts and Properties of Combinations of Subunits with ATPase Activity
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概要
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Subunits (α, β, γ) and mixtures of subunits (αβ, αγ, αβγ, αβyε) were isolated without denaturation from a chloroform extract of chloroplast coupling factor 1 (CF_1) from maize (Zea mays var. Ushiku 5-4) and from spinach by fast protein liquid chromatography (FPLC), on an anion-exchange column of Mono-Q in the presence of n-octylglucoside (OG) and on a chromatofocusing column of Mono-P. The α β γ-subunit complex (CF_1 α β y) was the minimum unit required for ATPase activity, as was confirmed by the reconstituted complex of α β and γ subunits. An ε subunit isolated from maize inhibited the ATPase activity of CF_1 α β γ from both maize and spinach. CF_1 α β γ was found to contain an OG-dependent Mg^<2+>-ATPase. The ATPase activity of CF_1 α β y required divalent cations, such as Mg^<2+> or Mn^<2+>, for its expression in the presence of OG; its optimum pH was 8.0 and it was markedly inhibited by NaN_3. The enzyme hydrolyzed ATP in prefernece to GTP but not CTP, UTP, ADP, AMP or pNPP. Lineweaver-Burk plots of its activity were curvilinear in the range of 0.6-0.7 mM ATP・Mg^<2+>.
- 日本植物生理学会の論文
著者
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Kasamo Kunihiro
Department of Applied Physiology, National Institute of Agrobiological Resources
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Arai Yasuhito
Department Of Cell Biology National Institute Of Agrobiological Resources:(present)department Of Bio
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Kagita Fumiko
Molecular Function Laboratory National Food Research Institute
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Kagita Fumiko
Department of Cell Biology, National Institute of Agrobiological Resources
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