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Structure function relationship of vanadate bound to a single site in chloroplast CF1-ATPase as determined by X-ray absorption

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Abstract

The structure of vanadate, a phosphate analogue which was suggested to function in the presence of tightly bound ADP and divalent cations as a transition state inhibitor of CF1-ATPase, was investigated by X-ray absorption spectroscopy. Analysis of the vanadium K-edge was used for determination of the structure of vanadate bound to a single site in CF1-ATPase containing a single tightly bound ADP. There was a decrease in the intensity of the 1s-3d pre-edge transition and a change in the shape of two other shoulders at the edge region upon binding of vanadate to CF1 in the presence of Mg2+ ions. The changes are due to alteration in the structure of vanadium from tetrahedral to a five-coordinated trigonal bipyramidal geometry. Comparison of the pre-edge peak intensity of ADP-vanadate complex, and model compound resolved by crystallography support the proposed structure of CF1-bound vanadate. 51V NMR measurements were used to verify the pentacoordinated structure of ADP-vanadate complex used as a model in the X-ray absorption studies. The inhibition of a single and multiple site activity by vanadate and by MgADP was measured. Vanadate inhibition of CF1-ATPase activity decreased more than 90 fold in the presence of MgADP. A differential specificity of the inhibition in single and multiple mode of activity was observed. It is suggested that ADP-vanadate binds to the active sites of the enzyme as a pentacoordinated vanadium having approximate trigonal bipyramidal geometry. This structure is analogous to the proposed transition state of the phosphate during the synthesis and the hydrolysis of ATP by CF1.

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Authors and Affiliations

  1. Department of Structural Biology, The Weizmann Institute for Science, The Netherlands

    Irit Sagi & Grant Bunker

  2. Biochemistry, The Netherlands

    Yehosua Hochman & Chanoch Carmeli

  3. Tel Aviv University, Tel Aviv, 69978, Israel

    Shmuel Carmeli

Authors
  1. Irit Sagi
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  2. Yehosua Hochman
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  3. Grant Bunker
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  4. Shmuel Carmeli
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  5. Chanoch Carmeli
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Sagi, I., Hochman, Y., Bunker, G. et al. Structure function relationship of vanadate bound to a single site in chloroplast CF1-ATPase as determined by X-ray absorption. Photosynthesis Research 57, 275–285 (1998). https://doi.org/10.1023/A:1006020813909

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  • DOI: https://doi.org/10.1023/A:1006020813909

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