Abstract
Hg2+ binding to ouabain-sensitive Na+-K+-ATPase of rat platelet membrane was specific with a Ka of 1.3×109 moles and Bmax of 3.8 nmoles/mg protein. The binding of mercury to Na+-K+-ATPase also inhibits the enzyme significantly (P<0.001), which is greater than its ouabain sensitivity. Further in the cytosol of washed platelets conjugation of reduced glutathione (GSH) to Hg2+ is correlated dose dependently (25, 50 and 100 pmoles) to enhanced GSH-S-transferase (GST) activity. It may be concluded from the present in vitro experiments that mercury binds specifically to thiol groups present in the platelet membrane Na+-K+-ATPase, inhibits the enzyme and induces changes in platelet function, namely, platelet aggregation by interfering with the sodium pump.
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References
Anner BM, Moosmayer M, Imesch E. 1990 Chelation of mercury by ouabain-sensitive and ouabain-resistant Na-K-ATPase. Biochem Biophys Res Commun 167, 115–1121.
Anner BM, Moosmayer M. 1992 Mercury inhibits Na-K-ATPase primarily at the cytoplasmic side. Am J Physiol 262, 843–848.
Ashida S, Abiko Y. 1979 Mode of action of ticlopidine in inhibition of platelet aggregation in the rat. Thromb Haemost 41, 436–449.
Bhattacharya Shelley, Bose S, Mukhopadhyay B et al. 1997 Specific binding of inorganic mercury to Na+-K+-ATPase in rat liver plasma membrane and signal transduction. BioMetals 10, 157–162.
Bhattacharya S. 1998 Mechanisms of signal transduction in the stress response of hepatocytes. In: Jeon KW, ed. International Review of cytology. New York: Academic Press; 109–156.
Bose S, Ghosh P, Ghosh S et al. 1993 Time dependent distribution of [203Hg] mercuric nitrate in the subcellular fractions of rat and fish liver. Biomed Environ Sci 6, 195–206.
Chaudhury S, Mukhopadhyay B, Bose S, Bhattacharya S. 1996 Involvement of mercury in platelet aggregation-probable mechanism of action. Biomed Environ Sci 9, 26–36.
Denizeau F, Marion M, Chtaib M, Schmit JP. 1990. Toxicity of heavy metals in cultured hepatocytes. Environ Toxicol Chem 9, 737–743.
Dorn GW. 1990 Cyclic oxidation-reduction reactions regulate thromboxane A2/prostaglandin H2 receptor number and affinity in human platelet membranes. J Biol Chem 265, 4240–4246.
Ellman GL, Courtney KD, Andres B Jr, Featherstone RM. 1961 A new and rapid colorimetric determination of acetylcholinesterase activity. Biochem Pharmacol 7, 88–95.
Feinberg H, Sandler WC, Scorer M, Le Breton GC, Grossman B, Born GVR. 1977 Movement of sodium into human platelets induced by ADP. Biochim Biophys Acta 470, 317–324.
Fiske CH, Subbarow Y. 1925 The colorimetric determination of phosphorus. J Biol Chem 66, 375–400.
Gennis RB. 1989 Biomembranes: Molecular structure and function. New York: Springer Verlag.
Ghosh N, Bhattacharya S. 1992. Thyrotoxicity of the chlorides of cadmium and mercury in rabbit. Biomed Environ Sci 5, 236–240.
Habig WH, Pabst MJ, Jakoby WB. 1974 Glutathione-S-transferase the first enzymatic step in mercapturic acid formation. J Biol Chem 249, 7130–7139.
Haslam RJ, Lynham JA. 1972 Activation and inhibition of blood platelet adenylate cyclase by adenosine or by 2-chloroadenosine. Life Sci 11, 1143–1154.
Hayashi Y, Takenaka S, Kohmura C, Ikeda H. 1998 Preparation of discoid washed platelets by differential centrifugation. Clinica Chimica Acta 275, 99–105.
Hughes WL. 1950 Protein mercaptides. Cold Spring Harbour Symposia on Quantitative Biology 14, 79–84.
Imesch E, Moosmayer M, Anner BM. 1992 Mercury weakens membrane anchoring of Na-K-ATPase. Amer J Physiol 262, 837–842.
Imura N, Miura K, Inokawa M, Inokawa M, Nakada S. 1980 Mechanisms of methylmercury cytotoxicity: By biochemical and morphological experiments using cultured cells. Toxicology 17, 241–254.
Inglis, JK. 1980 Animal health and hygiene. In Introduction to Laboratory Animal Science and Technology. Oxford: Pergamon Press; 37–66.
Jamaluddin M, Thomas A. 1992 Competitive inhibition of hydrogen peroxide-induced aggregation of calf platelets by prostaglandin H2/thromboxane A2 receptor ligands. J Biosci 17, 129–140.
Klonne DR, Johnson DR. 1998 Enzyme activity and sulfhydryl status in rat renal cortex following mercuric chloride and dithiothreitol administration. Toxicol Lett 42, 199–205.
Lanzetta PA, Alvarez LJ, Reinach PS, Candia OA. 1979 An improved assay for nanomole amounts of inorganic phosphate. Anal Biochem 100, 95–97.
Lionetto MG, Maffia M, Cappello MS, Giordano ME, Storelli C, Schettino T. 1998 Effect of cadmium on carbonic anhydrase and Na+-K+-ATPase in eel, Anguilla anguilla, intestine and gills. Comp Biochem Physiol 120, 89–91.
Lowry OH, Rosebrough VJ, Farr AL, Randall RJ. 1951 Protein measurement with the Folin-phenol reagent. J Biol Chem 193, 265–275.
Mondal S, Mukhopadhyay B, Bhattacharya S. 1997 Inorganic mercury binding to fish oocyte plasma membrane induces steroidogenesis and translatable messenger RNA synthesis. BioMetals 10, 1–6.
Mukhopadhyay S, Addya S, Bhattacharya DK, Chatterjee GC. 1988 Effects of cadmium treatment in vitro on the antioxidant protection mechanism and activation of human blood platelets. Thromb Res 50, 419–427.
Pederson PL, Carafoli E. 1987 Ion-motive ATPases 1. ubiquity, properties and significance to cell function. Trends in Biochem Sciences 12, 146–150.
Plummer DT. 1998 An Introduction to Practical Biochemistry. New Delhi: Tata McGraw-Hill Pub. Co. Ltd.
Rabenstein DL. 1978 The aqueous solution chemistry of methylmercury and its complexes. Accounts Chem Res 11, 100–107.
Rothstein A. 1959 Cell membrane as site of action of heavy metals. Fed Proc 18, 1026–1038.
Snedecor GW, Cochran WG. 1967 Student's t-distribution. In Statistical Methods. Ames: Iowa State University Press; 59–61.
Vinaya Kumar S, Bhattacharya S. 2000 in vitro toxicity of mercury, cadmium and arsenic to platelet aggregation: Influence of adenylate cyclase and phosphodiesterase activity. In vitro Molec Toxicol 13, 137–144.
Vinaya Kumar S, Bose R, Bhattacharya S. 2001 Low doses of heavy metals disrupt normal structure and function of rat platelets. J Env Pathol Toxicol Oncol 20, 69–79.
Webb JL. 1966 In Enzyme and Metabolic inhibitors. Vol. III, pp 595–793. New York: Academic Press.
Zbikowska HM, Wachowicz B, Krajewski T. 1997 Comparitive effects of selenite and selenate on the glutathione-related enzymes activity in pig blood platelets. Biol Trace Elem Res 57, 259–269.
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Kumar, S.V., Maitra, S. & Bhattacharya, S. In vitro binding of inorganic mercury to the plasma membrane of rat platelet affects Na+-K+-ATPase activity and platelet aggregation. Biometals 15, 51–57 (2002). https://doi.org/10.1023/A:1013196611595
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DOI: https://doi.org/10.1023/A:1013196611595