Ukr.Biochem.J. 2015; Volume 87, Issue 3, May-Jun, pp. 47-56

doi: https://doi.org/10.15407/ubj87.03.047

Carbonic anhydrase activity of integral-functional complexes of thylakoid membranes of spinach chloroplasts

08.07.2015   Uncategorized   No comments

A. V. Semenihin, O. K. Zolotareva

M. G. Kholodny Institute of Botany, National Academy
of Sciences of Ukraine, Kyiv;
e-mail: membrana@ukr.net

Isolated thylakoid membranes were disrupted by treatment with nonionic detergents digitonin or dodecyl maltoside. Solubilized polypeptide complexes were separated by native gel charge shift electrophoresis. The position of ATP-synthase complex and its isolated catalytic part (CF1) within gel was determined using the color reaction for ATPase activity. Due to the presence of cytochromes, the red band in unstained gels corresponded to the cytochrome b6f complex. Localization of the cytochrome b6f complex, ATP synthase and coupling CF1 in the native gel was confirmed by their subunit composition determined after SDS-electrophoretic analysis. Carbonic anhydrase (CA) activity in polypeptide zones of PS II, cytochrome b6f complex, and ATP-synthase CF1 was identified in native gels using indicator bromothymol blue. CA activity of isolated CF1 in solution was determined by infrared gas analysis as the rate of bicarbonate dehydration. The water-soluble acetazolamide, an inhibitor of CA, unlike lipophilic ethoxyzolamide inhibited CA activity of CF1. Thus, it was shown for the first time that ATP-synthase has a component which is capable of catalyzing the interconversion of forms of carbonic acid associated with proton exchange. The data obtained suggest the presence of multiple forms of carbonic anhydrase in the thylakoid membranes of spinach chloroplasts and confirm their involvement in the proton transfer to the ATP synthase.

Keywords: , , , ,

References:

  1. Danielsson R, Suorsa M, Paakkarinen V, Albertsson PA, Styring S, Aro EM, Mamedov F. Dimeric and monomeric organization of photosystem II. Distribution of five distinct complexes in the different domains of the thylakoid membrane. J Biol Chem. 2006 May 19;281(20):14241-9.  PubMed, CrossRef
  2. Nevo R, Charuvi D, Tsabari O, Reich Z. Composition, architecture and dynamics of the photosynthetic apparatus in higher plants. Plant J. 2012 Apr;70(1):157-76. Review. PubMed, CrossRef
  3. Dekker JP, Boekema EJ. Supramolecular organization of thylakoid membrane proteins in green plants. Biochim Biophys Acta. 2005 Jan 7;1706(1-2):12-39. Review. PubMed, CrossRef
  4. Avenson TJ, Kanazawa A, Cruz JA, Takizawa K, Ettinger WE, Kramer DM. Integrating the proton circuit into photosynthesis: progress and challenges. Plant Cell Environ. 2005;28(1):97-109. CrossRef
  5. Zolotareva EK. Protonic regulation of the process of photosynthetic energy transformation. Fiziol Biochim Kult Rast. 2010;42:37-50. (In Russian).
  6. Villarejo A, Shutova T, Moskvin O, Forssén M, Klimov VV, Samuelsson G. A photosystem II-associated carbonic anhydrase regulates the efficiency of photosynthetic oxygen evolution. EMBO J. 2002 Apr 15;21(8):1930-8. PubMed,  PubMedCentral, CrossRef
  7. Ignatova LK, Rudenko NN, Khristin MS, Ivanov BN. Heterogeneous nature of carbonic anhydrase activity of thylakoid membranes. Biochemistry (Mosc.). 2006;71(5):525-532. (In Russian). CrossRef
  8. Ignatova LK, Rudenko NN, Mudrik VA, Fedorchuk TP, Ivanov BN. Carbonic anhydrase activity in Arabidopsis thaliana thylakoid membrane and fragments enriched with PSI or PSII. Photosynth Res. 2011 Dec;110(2):89-98. PubMed, CrossRef
  9. Shutova T, Kenneweg H, Buchta J, Nikitina J, Terentyev V, Chernyshov S, Andersson B, Allakhverdiev SI, Klimov VV, Dau H, Junge W, Samuelsson G. The photosystem II-associated Cah3 in Chlamydomonas enhances the O2 evolution rate by proton removal. EMBO J. 2008 Mar 5;27(5):782-91.  PubMed, PubMedCentral, CrossRef
  10. Klimov VV, Hulsebosch RJ, Allakhverdiev SI, Wincencjusz H, van Gorkom HJ, Hoff AJ. Bicarbonate may Be required for ligation of manganese in the oxygen-evolving complex of photosystem II. Biochemistry. 1997 Dec 23;36(51):16277-81. PubMed, CrossRef
  11. Lu YK, Stemler AJ. Extrinsic photosystem II carbonic anhydrase in maize mesophyll chloroplasts. Plant Physiol. 2002 Feb;128(2):643-9. PubMed, PubMedCentral, CrossRef
  12. Burnell JN, Gibbs MJ, Mason JG. Spinach chloroplastic carbonic anhydrase: nucleotide sequence analysis of cDNA. Plant Physiol. 1990 Jan;92(1):37-40. PubMed, PubMedCentral, CrossRef
  13. Arnon DI. Copper enzymes in isolated chloroplasts. polyphenoloxidase in beta vulgaris. Plant Physiol. 1949 Jan;24(1):1-15. PubMed, PubMedCentral, CrossRef
  14. 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
  15. Andersson L, Borg H, Mikaelsson M. Molecular weight estimations of proteins by electrophoresis in polyacrylamide gels of graded porosity. FEBS Lett. 1972 Feb 1;20(2):199-202. PubMed, CrossRef
  16. Kolesnichenko AV, Ostroumova EA, Zykova VV, Voinikov VK. Proteins of four species of grasses, immunochemically related stress protein 310 kD. Plant Physiol. 2000;47:199-202. (In Russian).
  17. Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680-5. PubMed, CrossRef
  18. Allen JM, Hyncik G. Localization of alkaline phosphatases in gel matrices following electrophoresis. J Histochem Cytochem. 1963;11(2):169-175.  CrossRef
  19. Gomori G. Preparation of buffers for use in enzyme studies. Methods Enzymol. 1955;1:138-146. CrossRef
  20. Edwards LJ, Patton RL. Visualization of carbonic anhydrase activity in polyacrylamide gels. Stain Technol. 1966;41:303-304.
  21. Lien S, Racker E. Preparation and assay of chloroplast coupling factor CF1. Methods Enzymol. 1971;23:547-555. CrossRef
  22. Nikulina GI. Review of methods for the colorimetric determination of phosphorus on the formation of molybdenum blue. Moscow: Nauka,1965; 45 p.
  23. Järvi S, Suorsa M, Paakkarinen V, Aro EM. Optimized native gel systems for separation of thylakoid protein complexes: novel super- and mega-complexes. Biochem J. 2011 Oct 15;439(2):207-14. PubMed, CrossRef
  24. Shao J, Zhang Y, Yu J, Guo L, Ding Y. Isolation of Thylakoid Membrane Complexes from Rice by a New Double-Strips BN. SDS-PAGE and Bioinformatics Prediction of Stromal Ridge Subunits Interaction. PLoS ONE. 2011;6(5):e20342.  CrossRef
  25. 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
Creative CommonsThis work is licensed under a Creative Commons Attribution 4.0 International License.