Ukr.Biochem.J. 2017; Volume 89, Issue 4, Jul-Aug, pp. 43-48


Reversible pH-dependent activation/inactivation of CF(1)-ATPase of spinach chloroplasts

A. P. Khomochkin, O. B. Onoiko, A. V. Semenikhin, O. K. Zolotareva

M. G. Kholodhy Institute of Botany, National Academy of Sciences of Ukraine, Kyiv;

The aim of the work was to study the reverse pH-dependent regulation of the enzymatic activity of the catalytic part of ATP synthase (EC of chloroplast – coupling factor CF1. It was shown that the short-term incubation of isolated CF1 in the media with pH 4.5 or 3.5 leads to inactivation of Ca2+-ATPase, which is rapidly (t1/2 ~ 1 min) restored in the medium containing 0.5-10 mM bicarbonate at pH 7.8. After acid treatment, the rate of Mg2+-ATPase reaction was also stimulated in the presence of 1 mM bicarbonate (рН 7.8; 37 °С). The increase in Ca2+– and Mg2+-АТР activity of CF1 associated with the addition of NaHCO3 solution was completely eliminated after the introduction of 50 mM acetazolamide – a specific inhibitor of carbonic anhydrase. The obtained results suggest the existence of the bound bicarbonate in the CF1 structure, which apparently participates in proton transfer.

Keywords: , , , , ,


  1. Junge W, Nelson N. ATP synthase. Annu Rev Biochem. 2015;84:631-57. PubMed, CrossRef
  2. Malyan AN. Noncatalytic nucleotide binding sites: properties and mechanism of involvement in ATP synthase activity regulation. Biochemistry (Mosc). 2013 Dec;78(13):1512-23. PubMed, CrossRef
  3. Groth G, Pohl E. The structure of the chloroplast F1-ATPase at 3.2 A resolution. J Biol Chem. 2001 Jan 12;276(2):1345-52. PubMed, CrossRef
  4. Tiedge H, Lünsdorf H, Schäfer G, Schairer HU. Subunit stoichiometry and juxtaposition of the photosynthetic coupling factor 1: Immunoelectron microscopy using monoclonal antibodies. Proc Natl Acad Sci USA. 1985 Dec;82(23):7874-8. PubMed, PubMedCentral, CrossRef
  5. Bakker-Grunwald T, van Dam K. On the mechanism of activation of the ATPase in chloroplasts. Biochim Biophys Acta. 1974 May 22;347(2):290-8. PubMed, CrossRef
  6. Malyan AN, Vitseva OI, Gubanova ON. Mg2+-dependent inactivation/H+-dependent activation equilibrium of chloroplast F1-ATPase. Photosynth Res. 1998; 57(3): 297-303. CrossRef
  7. Zolotareva EK, Gasparyan ME, Yaguzhinsky LS. Transfer of tightly-bound tritium from the chloroplast membranes to CF1 is activated by the photophosphorylation process. FEBS Lett. 1990 Oct 15;272(1-2):184-6. PubMed, CrossRef
  8. Malyan AN. A possible mechanism of coupling ATP synthesis with proton transfer in the enzymatic complex of chloroplast CF1. Rep Acad Sci USSR. 1986; 291: 1015-1018. (In Russian).
  9. Zolotareva EK, Tereshchenko AF, Dovbysh EF, Onoiko EB. Effect of alcohols on inhibition of photophosphorylation and electron transport by N,N’-dicyclohexylcarbodiimide in pea chloroplasts. Biochemistry (Moscow). 1997; 62(6): 631-635.
  10. Zakharov SD, Sytnik SK, Mal’yan AN, Makarov AD. Isolation and properties of CF1 ATPase chloroplasts with changed submolecular structure. Biokhimiia. 1978 May;43(5):887-91. (In Russian). PubMed
  11. Arnon DI. Copper enzymes in isolated chloroplasts. Polyphenoloxidase in Beta vulgaris. Plant Physiol. 1949 Jan;24(1):1-15. PubMed, PubMedCentral, CrossRef
  12. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265-75. PubMed
  13. Semenihin AV, Zolotareva EK. Identification of carbonic anhydrase activity associated with protein complexes of photosynthetic membranes of spinach chloroplasts. Rep Nat Acad Sci Ukr. 2014; 9: 141-145. (In Ukrainian).
  14. Nikulina GN. A review of the methods for the colorimetric determination of phosphorus by the formation of molybdenum blue. М.-L.: Nauka, 1965. 45 p. (In Russian).
  15. Murataliev MB, Boyer PD. The mechanism of stimulation of MgATPase activity of chloroplast F1-ATPase by non-catalytic adenine-nucleotide binding. Acceleration of the ATP-dependent release of inhibitory ADP from a catalytic site. Eur J Biochem. 1992 Oct 15;209(2):681-7. PubMed, CrossRef
  16. Guerrero KJ, Xue ZX, Boyer PD. Active/inactive state transitions of the chloroplast F1 ATPase are induced by a slow binding and release of Mg2+. Relationship to catalysis and control of F1 ATPases. J Biol Chem. 1990 Sep 25;265(27):16280-7. PubMed
  17. Onoiko E. B., Polishchuk A. V., Zolotareva E. K. Stimulation of photophosphorylation in spinach isolated chloroplasts by exogenous bicarbonate: role of carbonic anhydrase. Rep Nat Acad Sci Ukr. 2010; 10: 160-165. (In Russian).
  18. van Rensen JJ. Role of bicarbonate at the acceptor side of Photosystem II. Photosynth Res. 2002;73(1-3):185-92. PubMed, CrossRef
  19. Semenihin AV, Zolotareva OK. Carbonic anhydrase activity of integral-functional complexes of thylakoid membranes of spinach chloroplasts. Ukr Biochem J. 2015 May-Jun;87(3):47-56. PubMed, CrossRef
  20. Fabre N, Reiter IM, Becuwe-Linka N, Genty B, Rumeau D. Characterization and expression analysis of genes encoding alpha and beta carbonic anhydrases in Arabidopsis. Plant Cell Environ. 2007 May;30(5):617-29. PubMed, CrossRef
  21. Rudenko NN, Ignatova LK, Fedorchuk TP, Ivanov BN. Carbonic anhydrases in photosynthetic cells of higher plants. Biochemistry (Mosc). 2015 Jun;80(6):674-87. PubMed, CrossRef

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