Advertisement

Effect of phosphate on the kinetics and specificity of glycation of protein.

Open AccessPublished:May 25, 1987DOI:https://doi.org/10.1016/S0021-9258(18)48224-5
      This paper is only available as a PDF. To read, Please Download here.
      The glycation (nonenzymatic glycosylation) of several proteins was studied in various buffers in order to assess the effects of buffering ions on the kinetics and specificity of glycation of protein. Incubation of RNase with glucose in phosphate buffer resulted in inactivation of the enzyme because of preferential modification of lysine residues in or near the active site. In contrast, in the cationic buffers, 3-(N-morpholino)propane-sulfonic acid and 3-(N-tris(hydroxymethyl)methyl-amino)-2-hydroxypropanesulfonic acid, the kinetics of glycation of RNase were decreased 2- to 3-fold, there was a decrease in glycation of active site versus peripheral lysines, and the enzyme was resistant to inactivation by glucose. The extent of Schiff base formation on RNAse was comparable in the three buffers, suggesting that phosphate, bound in the active site of RNase, catalyzed the Amadori rearrangement at active site lysines, leading to the enhanced rate of inactivation of the enzyme. Phosphate catalysis of glycation was concentration-dependent and could be mimicked by arsenate. Phosphate also stimulated the rate of glycation of other proteins, such as lysozyme, cytochrome c, albumin, and hemoglobin. As with RNase, phosphate affected the specificity of glycation of hemoglobin, resulting in increased glycation of amino-terminal valine versus intrachain lysine residues. 2,3-Diphosphoglycerate exerted similar effects on the glycation of hemoglobin, suggesting that inorganic and organic phosphates may play an important role in determining the kinetics and specificity of glycation of hemoglobin in the red cell. Overall, these studies establish that buffering ions or ligands can exert significant effects on the kinetics and specificity of glycation of proteins.

      REFERENCES

        • Thorpe S.R.
        • Baynes J.W.
        Horowitz M.I. The Glycoconjugates. 3. Academic Press, New York1982: 113-132
        • Isbell H.S.
        • Frush H.
        J. Org. Chem. 1958; 23: 1309-1319
        • Gottschalk A.
        Gottschalk A. The Glycoproteins. Elsevier/North Holland, New York1972: 141-157 (Part A)
        • Garner M.H.
        • Bogart Jr., R.A.
        • Gurd F.R.N.
        J. Biol. Chem. 1975; 250: 4398-4404
        • Shapiro R.
        • McManus M.J.
        • Zalut C.
        • Bunn H.F.
        J. Biol. Chem. 1980; 255: 3120-3127
        • Watkins N.G.
        • Thorpe S.R.
        • Baynes J.W.
        J. Biol Chem. 1985; 260: 10629-10636
        • Garlick R.L.
        • Mazer J.S.
        J. Biol. Chem. 1983; 258: 6142-6146
        • Iberg N.
        • Flückiger R.
        J. Biol. Chem. 1986; 261: 13542-13545
        • Baynes J.W.
        • Ahmed M.U.
        • Fisher C.I.
        • Hull C.J.
        • Lehman T.A.
        • Watkins N.G.
        • Thorpe S.R.
        Dev. Fd. Sci. 1986; 13: 421-431
        • Schwartz B.A.
        • Gray G.R.
        Arch. Biochem. Biophys. 1977; 181: 542-549
        • Baynes J.W.
        • Thorpe S.R.
        • Murtiashaw M.H.
        Methods Enzymol. 1984; 106: 88-98
        • Bissé E.
        • Berger W.
        • Flückiger R.
        Diabetes. 1982; 31: 630-633
        • Neglia C.I.
        • Cohen H.J.
        • Garber A.R.
        • Ellis P.D.
        • Thorpe S.R.
        • Baynes J.W.
        J. Biol. Chem. 1983; 258: 14279-14283
        • Neglia C.I.
        • Cohen H.J.
        • Garber A.R.
        • Thorpe S.R.
        • Baynes J.W.
        J. Biol. Chem. 1985; 260: 5406-5410
        • Antonini E.
        • Brunori M.
        Hemoglobin and Myoglobin and Their Reactions with Ligands. Elsevier Scientific Publishing Co., Amsterdam1971
        • Clegg J.B.
        • Naughton M.A.
        • Weatherall D.J.
        J. Mol. Biol. 1966; 19: 91-108
        • Good N.E.
        • Winget G.D.
        • Winter W.
        • Connolly T.N.
        • Izawa S.
        • Singh R.M.M.
        Biochemistry. 1966; 5: 467-473
        • Ferguson W.J.
        • Braunschweiger K.I.
        • Smith J.R.
        • McCormick J.J.
        • Wasmann C.C.
        • Jarvis N.P.
        • Bell D.H.
        • Good N.E.
        Anal. Biochem. 1980; 104: 300-310
        • Baynes J.W.
        • Wold F.
        J. Biol. Chem. 1976; 251: 6016-6024
        • Richards F.M.
        • Wyckoff H.W.
        Boyer P.D. The Enzymes. 3rd Ed. IV. Academic Press, New York1971: 648-806
        • Beutler E.
        • Srivastava S.K.
        Williams W.J. Beutler E. Erslev A.J. Rundles R.W. Hematology. McGraw-Hill Book Co., New York1972: 94-100
        • Reynolds T.M.
        Adv. Food Res. 1963; 12: 1-52
        • Reynolds T.M.
        Adv. Food Res. 1965; 14: 167-283
        • Webb B.H.
        J. Dairy Sci. 1935; 2: 81-96
        • Kato H.
        Bull. Agr. Chem. Soc. Japan. 1956; 20: 273-278
        • Burton H.S.
        • McWeeny D.J.
        Nature. 1963; 197: 266-268
        • Thornalley P.
        • Wolff S.
        • Crabbe J.
        • Stern A.
        Biochim. Biophys. Acta. 1984; 797: 276-287
        • Dickerson R.E.
        • Geis I.
        The Structure and Action of Proteins. W. H. Benjamin, Inc., Menlo Park, CA1969
        • Smith R.J.
        • Koenig R.J.
        • Binnerts A.
        • Soeldner J.S.
        • Aoki T.T.
        J. Clin. Invest. 1982; 69: 1164-1168
        • Roberts A.P.
        • Story C.J.
        • Ryall R.G.
        Diabetologia. 1984; 26: 389-391
        • Lowrey C.H.
        • Lyness S.J.
        • Soeldner J.S.
        J. Biol. Chem. 1985; 260: 11611-11618
        • Bunn H.F.
        • Shapiro R.
        • McManus M.
        • Garrick L.
        • McDonald M.J.
        • Gallop P.M.
        • Gabbay K.H.
        J. Biol. Chem. 1979; 254: 3892-3898
        • Hedlund B.
        • Danielson C.
        • Lovrien R.
        Biochemistry. 1972; 11: 4660-4668
        • Acharya A.S.
        • Manning J.M.
        J. Biol. Chem. 1980; 255: 1406-1412
        • Acharya A.S.
        • Sussman L.G.
        • Manning J.M.
        J. Biol. Chem. 1983; 258: 2296-2302