Purification And Characterization Of A Low-molecular-weight Bovine Kidney Acid Phosphatase

A relative low molecular mass bovine kidney acid phosphatase was purified 1,640-fold to homogeneity, with 7% recovery. The purified enzyme (specific activity 100 μmol min-1 mg-1) was electrophoretically homogeneous with a relative molecular mass of 17.8 kDa, as determined by SDS-polyacrylamide gel electrophoresis. A broad pH optimum of 4.0-5.5 and a maximal enzyme activity at 60°C were determined for the p-nitrophenyl phosphate hydrolysis. Apparent Km values of 0.14 mM, 0.4 mM, 0.3 mM and 7.9 mM were obtained, at 37°C and pH 5.0, for the best substrates p-nitrophenyl phosphate, β-naphtyl-phosphate, flavin mononucleotide and tyrosine-phosphate, respectively. The enzyme activity was enhanced by guanosine but inhibited by ZnCl2 and CuSO4, p-cloromercuribenzoate and ammonium molybdate. Vanadate (Ki 0.47 μM), pyridoxal 5′-phosphate (Ki 2.2 μM), inorganic phosphate (Ki 0.77 mM) are competitive inhibitors. Both glycerol and methanol increased significantly the acid phosphatase activity, acting as good phosphate acceptors in the transphosphorylation reaction.694451460Araújo, P.S., Mies, V., Miranda, O., Subcelular distribution of low-molecular-weight acid phosphatases (1976) Biochim. Biophys. Acta, 452, pp. 121-130Baldijão, C.E.M., Guija, E., Bittencourt, H.M.S., Chaimovich, H., Steady state kinetics and effects of inhibitors on acid phosphatase from bovine brain (1975) Biochim. Biophys. Acta, 391, pp. 316-325Bhargava, R., Sachar, R.C., Induction of acid phosphatase in cotton seedlings enzyme purification, subunit structure and kinetic properties (1987) Phytochemistry, 26, pp. 1293-1297Baxter, J.H., Suelter, C.H., Resolution of the low-molecular-weight acid phosphatase in avian pectoral muscle into distinct enzyme forms (1985) Arch. Biochem. Biophys., 239, pp. 29-37Berti, A., Rigacci, S., Raugei, G., Degl'innocenti, D., Ramponi, G., Inhibition of cellular esponse to platelet-derived growth factor by low Mr phosphotyrosine protein phosphatase overexpression (1994) FEBS Lett., 349, pp. 7-12Camici, G., Manao, G., Cappugi, G., Modesti, A., Stefani, M., Ramponi, G., The complete amino acid sequence of the low molecular weight cytosolic acid phosphatase (1989) J. Biol. Chem., 264, pp. 2560-2567Chaimovich, H., Nome, F., Purification and properties of an acid phosphatase from bovine brain (1970) Arch. Biochem. Biophys., 139, pp. 9-16Chernoff, J., Li, H.C., A major phosphotyrosyl-protein phosphatase from bovine heart is associated with a low-molecular-weight acid phosphatase (1985) Arch. Biochem. Biophys., 240, pp. 135-145Chiarugi, P., Marzocchini, R., Raugei, G., Pazzagli, C., Berti, A., Camici, G., Manao, G., Ramponi, G., Differential role of four cysteines residues on the activity of a low Mr phosphotyrosine protein phosphatase (1992) FEBS Lett., 310, pp. 9-12Chiarugi, P., Cirri, P., Camici, G., Manao, G., Fiaschi, T., Raugei, G., Cappugi, G., Ramponi, G., The role of His66 and His72 in the reaction mechanism of bovine low-Mr phosphotyrosine protein phosphatase (1994) Biochem. J., 298, pp. 427-433Chiarugi, P., Cirri, P., Raugei, G., Camici, G., Dolfi, F., Berti, A., Ramponi, G., PDGF receptor as a specific in vivo target for low Mr phosphotyrosine protein phosphatase (1995) FEBS Lett., 372, pp. 49-53Davis, J.P., Zhou, M.-M., Van Etten, R.L., Kinetic and site directed mutagenesis studies of the cysteine residues of bovine low molecular weight phosphotyrosyl protein phosphatase (1994) J. Biol. Chem., 269, pp. 8734-8740De-Kundu, P., Banerjee, A.C., Multiple forms of acid phosphatase from seedlings axes of Vigna radiata (1990) Phytochemistry, 29, pp. 2825-2828Dissing, J., Svensmark, O., Human red cell acid phosphatase: Purification and properties of the A, B and C isoenzymes (1990) Biochim. Biophys. Acta, 1041, pp. 232-242Dissing, J., Johnsen, A.H., Human red cell acid phosphatase (ZCP1): The primary structure of two pairs of isoenzymes encoded by the ACP1*A and ACP1*C alleles (1992) Biochim. Biophys. Acta, 1121, pp. 261-268Dissing, J., Rangaard, B., Christensen, U., Activity modulation of the fas and slow isoenzymes of human cytosolic low-molecular-weight acid phosphatase (ACP1) by purines (1993) Biochim. Biophys. Acta, 1162, pp. 275-282Dixon, M., Webb, E.C., (1979) Enzymes, 3rd Edn, p. 1116. , Academic Press, New YorkDuff, S.M.G., Sarath, G., Plaxton, W.C., The role of acid phosphatases in plant phosphorus metabolism (1994) Physiol. Plant., 90, pp. 791-800Fauman, E.B., Saper, M.A., Structure and function of the protein tyrosine phosphatase (1996) Trends Biochem. Sci., 21, pp. 413-417Fuchs, K.R., Shekels, L.L., Bernlohr, D.A., Analysis of the ACP1 gene product: Classification as an FMN phosphatase (1992) Biochem. Biophys. Res. Commun., 189, pp. 1598-1605Fujimoto, S., Urata, Y., Nakagawa, T., Ohara, A., Characterization of intermediate-molecular-weight acid phosphatase from bovine kidney cortex (1984) J. Biochem., 96, pp. 1079-1088Galka, M., Dziembor-Gryszkiewicz, E., Kos, S., Ostrowski, W., Properties of low-molecular-weight acid phosphatases isolated from cytosol and chromatin of rat liver (1980) Acta Biochim. Pol., 27, pp. 281-293Hollander, V.P., Acid phosphatases (1971) The Enzymes, pp. 449-455. , BOYER, P. D. ed. New York, Academic PressKlarlund, J.K., Transformation of cells by an inhibitor of phosphatases acting on phosphotyrosine in proteins (1985) Cell, 41, pp. 707-717Kumagai, A., Dunphy, W.G., The cdc25 protein controls tyrosine dephosphorylation of the cdc2 protein in a cell-free system (1991) Cell, 64, pp. 903-914Logan, T.M., Zhou, M.-M., Nettesheim, D.G., Meadows, R.P., Van Etten, R.L., Fesik, S.W., Solution structure of a low molecular weight protein tyrosine phosphatase (1994) Biochemistry, 33, pp. 11087-11096Lowry, O.H., Lopez, J., The determination of inorganic phosphate in the presence of labile phosphate esters (1946) J. Biol. Chem., 162, pp. 421-424Lowry, O.H., Rosebrough, N., Farr, A., Randal, R., Protein measurement with the phenol reagent (1951) J. Biol. Chem., 193, pp. 265-275Panara, F., Angiolillo, A., Fagotti, A., Di Rosa, I., Francesca, S., Pascolini, R., Acid phosphatases in mammalian tissues. Evidence for the existence of a 57 kDa Zn2+-dependent acid phosphatase form (1992) Int. J. Biochem., 24, pp. 1619-1623Pot, D.A., Dixon, J.E., A thousand and two protein tyrosine phosphatases (1992) Biochim. Biophys. Acta, 1136, pp. 35-43Ramponi, G., Manao, G., Camici, G., Cappugi, G., Ruggiero, M., Bottaro, G.P., The 18 kDa cytosolic acid phosphatase from bovine liver has phosphotyrosine activity on the autophosphorylated epidermal growth factor receptor (1989) FEBS Lett., 250, pp. 469-473Reisfeld, R.A., Lewis, V.J., Williams, D.E., Disk electrophoresis of basic proteins and peptides on polyacrylamide gels (1962) Nature, 195, pp. 281-283Saeed, A., Tremori, E., Manao, G., Camici, G., Cappugi, G., Ramponi, G., Bovine brain low Mr acid phosphatase: Purification and properties (1990) Physiol. Chem. Phys. and Med. NMR, 22, pp. 81-94Su, X.-D., Taddei, N., Stefani, M., Ramponi, G., Nordlund, P., The crystal structure of a low-molecular-weight phosphotyrosine protein phosphatase (1994) Nature, 370, pp. 575-578Taga, E.M., Van Etten, R.L., Human liver acid phosphatase: Purification and properties of a low molecular weight isoenzymes (1982) Arch. Biochem. Biophys., 214, pp. 505-515Tantzaki, M.M., Bittencourt, H.M.S., Chaimovich, H., Activation of low molecular weight acid phosphatase from bovine brain by purines and glycerol (1976) Biochim. Biophys. Acta, 485, pp. 116-123Ullah, A.H.J., Gibson, D.M., Purification and characterization of acid phosphatase from cotyledons of germinating soybean seeds (1988) Arch. Biochem. Biophys., 260, pp. 514-520Waheed, A., Laidler, P.M., Wo, Y.-Y.P., Van Etten, R.L., Purification and physicochemical characterization of a human placental acid phosphatase activity (1988) Biochemistry, 27, pp. 4265-4273Weber, K., Osborn, M., The reability of molecular weight determination by sodium dodecyl sulfate-polyacrilamide gel electrophoresys (1969) J. Biol. Chem., 244, pp. 4406-4412Zhang, Z.-Y., Van Etten, R.L., Purification and characterization of a low-molecular-weight acid phosphatase - A phosphotyrosyl-protein phosphatase from bovine heart (1990) Arch. Biochem. Biophys., 282, pp. 39-4