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Redox modulation of reductase and phosphatase activities in human erythrocytes

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We have previously reported that ferricyanide reductase activity in human erythrocytes depended on glycolysis and could be modulated by several compounds including oxidants and hormones like insulin. Insulin could activate glycolysis, probably as a consequence of tyrosine phosphorylation of protein band 3, implicating phosphorylation reactions as an important signal for activation of the reductase by insulin. Reversible phosphorylation of cellular proteins is also believed to play a key role in the action of insulin. Cytosolic acid phosphatase activity has been found in human erythrocytes. To further extend initial reports, we studied the effect of modulators on the cytosolic erythrocyte acid phosphatase. Mild oxidants like ferricyanide (1 mM), vanadate (1 mM), Mn2+ (0.5 and 1 mM), and phenylarsine oxide (10 and 100 μM) inhibited the phosphatase activity. Similarly, insulin at concentrations that stimulate ferricyanide reduction (500, 1000 μIU/ml) inhibited the activity of the phosphatase enzyme. The overall results indicated that oxidants are able to inhibit the acid phosphatase and stimulate the redox enzyme. In addition, a significant negative correlation (r = −0.400; P = 0.006) was observed between phosphatase and reductase activities. The observations discussed here, together with previous ones, emphasize that a close association between reductase and phosphatase enzymes may exist and also suggest a role for redox reactions in tyrosine phosphorylation/dephosphorylation-mediated signal transduction pathways.

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References

  • Benabe JE, Echegoyen LA, Pastrana B, Martinéz-Maldonado M (1987) Mechanism of inhibition of glycolysis by vanadate. J Biol Chem 262: 9555–9560

    Google Scholar 

  • Bernier M, Laird DM, Lane MD (1988) Effect of vanadate on the cellular accumulation of pp 15, an apparent product of insulin receptor tyrosine kinase action. J Biol Chem 263: 13626–13634

    Google Scholar 

  • Board PG (1981) NADH-ferricyanide reductase, a convenient approach to the evaluation of NADH-methaemoglobin reductase in human erythrocytes. Clin Chim Acta 109: 233–237

    Google Scholar 

  • Boivin P (1988) Role of the phosphorylation of red blood cell membrane proteins. Biochem J 256: 689–695

    Google Scholar 

  • —, Galand C (1986) The human red cell acid phosphatase is a phosphotyrosine protein phosphatase which dephosphorylates the membrane protein band 3. Biochem Biophys Res Commun 134: 557–564

    Google Scholar 

  • Bottini E, Modiano G, Santolamazza C, Filippi G, Bosinco L (1971) Studies of the in vitro effects of oxidized glutathione and acetylphenylhydrazine on acid phosphatase of human red blood cells: an experimental model for the investigation of hemolytic drug action at the molecular level. Clin Chim Acta 31: 243–254

    Google Scholar 

  • —, Bottini FG, Borgiani P (1997) Association between ACP1 and favism: a possible biochemical mechanism. Blood 89: 2613–2615

    Google Scholar 

  • Chernoff J, Li H-C (1985) A major phosphotyrosyl-protein phosphatase from bovine heart is associated with a low-molecularweight acid phosphatase. Arch Biochem Biophys 240: 135–145

    Google Scholar 

  • Clari G, Brunati AM, Moret V (1986) Partial purification and characterization of phosphotyrosyl-protein phosphatase(s) from human erythrocyte cytosol. Biochem Biophys Res Commun 137: 566–572

    Google Scholar 

  • —, Libera LD, Moret V (1990) Tyrosine phosphorylation of cytosolic proteins in human erythrocytes. Biochem Biophys Res Commun 166: 1378–1383

    Google Scholar 

  • —, Bordin L, Marzaro G, Moret V (1991) Effect of intracellular pH changes on the distribution of tyrosine- and serine/threonineprotein kinase activities in human erythrocytes. Biochem Biophys Res Commun 178: 1021–1027

    Google Scholar 

  • Cohen F (1989) The structure and regulation of protein phosphatases. Annu Rev Biochem 58: 453–508

    Google Scholar 

  • Conde M, Chiara, MD, Pintado E, Sobrino F (1995) Modulation of phorbol ester-induced respiratory burst by vanadate, genistein, and phenylarsine oxide in mouse macrophages. Free Rad Biol Med 18: 343–348

    Google Scholar 

  • Crane FL, Sun IL, Löwe HE (1990) Oxidoreductase enzymes in the plasma membrane. In: Crane FL, Morré DJ, Löw HE (eds) Oxidoreduction at the plasma membrane: relation to growth and transport, vol 1. CRC Press, Boca Raton, pp 29–65

    Google Scholar 

  • Dissing J, Dabl O, Svensmark O (1979) Phosphonic and arsonic acids as inhibitors of human red cell acid phosphatase and their use in affinity chromatography. Biochim Biophys Acta 569: 159–176

    Google Scholar 

  • —, Johnsen AH, Sensabaugh GF (1991) Human red cell acid phosphatase (ACPI). J Biol Chem 266: 20619–20625

    Google Scholar 

  • Gorin Y, Leseney AM, Ohayon R, Dupuy C, Pommier J, Virion A, Dème D (1997) Regulation of the thyroid NADPH-dependent H2O2 generator by Ca2+: studies with phenylarsine oxide in thyroid plasma membrane. Biochem J 321: 383–388

    Google Scholar 

  • Haring H-U, Kasuga M, White MF, Crettaz M, Kanh R (1984) Phosphorylation and dephosphorylation of the insulin receptor: evidence against an intrinsic phosphatase activity. Biochemistry 23: 3298–3306

    Google Scholar 

  • Harrison ML, Rathinavelu P, Arese P, Geahlen RL, Low PS (1991) Role of band 3 tyrosine phosphorylation in the regulation of erythrocyte glycolysis. J Biol Chem 266: 4106–4111

    Google Scholar 

  • Hecht D, Zick Y (1992) Selective inhibition of protein tyrosine phosphatase activities by H2O2 and vanadate in vitro. Biochem Biophys Res Commun 188: 773–779

    Google Scholar 

  • Heffetz D, Bushkin I, Dror R, Zick Y (1990) The insulinomimetic agents H2O2 and vanadate stimulate protein tyrosine phosphorylation in intact cells. J Biol Chem 265: 2896–2902

    Google Scholar 

  • Ide R, Maegawa H, Kikkawa R, Shigeta Y, Kashiwagi A (1994) High glucose condition activates protein tyrosine phosphatases and deactivates insulin receptor function in insulin-sensitive rat 1 fibroblasts. Biochem Biophys Res Commun 201: 71–77

    Google Scholar 

  • Li J, Elberg G, Shechter Y (1996) Phenylarsine oxide and vanadate: apparent paradox of inhibition of protein phosphotyrosine phosphatases in rat adipocytes. Biochim Biophys Acta 1312: 223–230

    Google Scholar 

  • Lucaciu CM, Dragu C, Copaescu L, Morariu V (1997) Manganese transport through human erythrocyte membranes: an EPR study. Biochim Biophys Acta 1328: 90–98

    Google Scholar 

  • Marques F, Crespo ME, Bicho M (1995) Control of NADH ferricyanide reductase activity in the human erythrocytes by somatotrophin and insulin. Redox Rep 1: 113–117

    Google Scholar 

  • — —, Pantaleão O, Bicho M (1996) Insulin activation of NADH ferricyanide reductase in human erythrocytes is mediated by the insulin receptor tyrosine kinase: a comparative study in normal and diabetic states. Redox Rep 2: 373–378

    Google Scholar 

  • Monteiro HP, Stern A (1996) Redox modulation of tyrosine phosphorylation dependent signal transduction pathways. Free Rad Biol Med 21: 323–333

    Google Scholar 

  • Stefani M, Caselli A, Bucciantini M, Pazzagli L, Dolfi F, Camici G, Manao G, Ramponi G (1993) Dephosphorylation of tyrosine phosphorylated synthetic peptides by rat liver phosphotyrosine protein phosphatase isoenzymes. FEBS Lett 326: 131–134

    Google Scholar 

  • Suzuki YJ, Forman HJ, Sevanian A (1997) Oxidants as stimulators of signal transduction. Free Rad Biol Med 22: 269–285

    Google Scholar 

  • Tuy FPD, Henry J, Rosenfeld C, Kahn A (1983) High tyrosine kinase activity in normal nonproliferating cells. Nature 305: 435–438

    Google Scholar 

  • — —, Kahn A (1985) Characterization of human red blood cell tyrosine kinase. Biochem Biophys Res Commun 126: 304–312

    Google Scholar 

  • Usui H, Kinohara N, Yoshikawa K, Imazu M, Imaoka T, Takeda M (1983) Phosphoprotein phosphatases in human erythrocyte cytosol. J Biol Chem 258: 10455–10463

    Google Scholar 

  • Valentine WN, Tanaka KR, Fredricks RE (1999) Erythrocyte acid phosphatase in health and disease. Am J Clin Pathol 36: 328–332

    Google Scholar 

  • van Kampen EJ, Zijlstra WG (1999) Standardization of hemoglobinometry: the hemoglobincyanide method. Clin Chim Acta 6: 538–544

    Google Scholar 

  • Wo YP, McCormack AL, Shabanowitz J, Hunt DF, Davis JP, Mitchell GL, van Etten RL (1992) Sequencing, cloning, and expression of human red cell-type acid phosphatase, a cytoplasmic phosphotyrosyl protein phosphatase. J Biol Chem 267: 10856–10865

    Google Scholar 

  • Zang W-R, Goldstein BJ (1991) Identification of skeletal muscle protein-tyrosine phosphatases by amplification of conserved cDNA sequences. Biochem Biophys Res Commun 178: 1291–1297

    Google Scholar 

  • Zipser Y, Kosower NS (1996) Phosphotyrosine phosphatase associated with band 3 protein in the human erythrocyte membrane. Biochem J 314: 881–887

    Google Scholar 

  • —, Piade A, Kosower NS (1997) Erythrocyte thiol status regulates band 3 phosphotyrosine level via oxidation/reduction of band 3- associated phosphotyrosine phosphatase. FEBS Lett 406: 126–130

    Google Scholar 

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Authors and Affiliations

  1. Departamento de Radioisótopos, Instituto de Técnicas Nucleares, Lisbon

    F. Marques

  2. Laboratório de Endocrinologia, Institute Portuguěs Oncologia de Francisco Gentil, Lisbon

    M. E. Crespo

  3. Laboratório de Genética, Faculdade de Medicina, Universidade de Lisboa, Av. Professor Egas Moniz, P-1699, Lisboa, Portugal

    M. E. Crespo, Z. I. Silva & M. Bicho

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  1. F. Marques
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  2. M. E. Crespo
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  3. Z. I. Silva
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  4. M. Bicho
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Marques, F., Crespo, M.E., Silva, Z.I. et al. Redox modulation of reductase and phosphatase activities in human erythrocytes. Protoplasma 206, 168–173 (1999). https://doi.org/10.1007/BF01279264

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  • DOI: https://doi.org/10.1007/BF01279264

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