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Ethanol induced alterations in plasma membrane protein phosphorylation of neurons and astrocytes

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Abstract

The endogenous protein phosphorylation patterns in plasma membranes of bulk isolated neurons and astroglia from control and chronic ethanol treated rats have been investigated. Chronic ethanol treatment resulted in increased phosphorylation of specific proteins with molecular weights 116, 63 and 60 kDa in both neurons and astrocytes. These proteins were further resolved by 2-DE and the analysis suggested an increased phosphorylation of ≏10–15 proteins, of which 116 kDa protein is phosphorylated to a higher extent by ethanol. Further, decreased phosphorylation was noticed in D-95 and D-63 proteins in neurons and D-78 and D-54 proteins in astrocytes. Alkali stability experiments for identifying the phosphoamino acid involved in phosphorylation of 116, 63 and 60 kDa proteins suggested that tyrosine and threonine are not involved and probably serine is the likely site for phosphorylation during chronic ethanol treatment. The phosphorylation of specific membrane proteins during chronic ethanol treatment might contribute to ethanol evoked cellular dysfunction.

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References

  1. Horvath TB: Clinical spectrum and epidemiological features of alcohol dementia. In: JG Rankin (ed.) Alcohol, Drugs and Brain Damage. Addiction Res Found Ontario, 1975, pp 1–16

  2. Goldstein DB: Alcohol and cellular membranes. In: U Rydberg, C Alling, J Engel (eds.) Alcohol and the Developing Brain. Raven Press, New York, 1985, pp 19–26

    Google Scholar 

  3. Chin JH, Goldstein DB: Effects of low concentrations of ethanol on the fluidity of spin labelled erythrocyte and brain membranes. Mol Pharmacol 13: 435–441, 1977

    PubMed  Google Scholar 

  4. Prakasa Babu P, Ratnakumari L, Vemuri MC: Differential changes in cell morphology, macromolecular composition and membrane protein profiles of neurons and astrocytes in chronic ethanol treated rats. Molecular and Cellular Biochemistry 130: 29–40, 1994

    PubMed  Google Scholar 

  5. Hidaka H, Hagiwara M, Chijiwa T: Molecular pharmacology of protein kinases. Neurochemical Res 15: 431–434, 1990

    Google Scholar 

  6. Nishizuka Y: The molecular heterogenity of protein kinase C and its implications for cellular regulation. Nature 334: 661–665, 1989

    Google Scholar 

  7. Matzel LD, Lederhendler II, Alkon DL: Regulation of short-term associative memory by calcium dependent protein kinase. J Neuroscience 10: 2300–2307, 1990

    Google Scholar 

  8. Hindmarch I, Kerr JS, Sherwood N: The effects of alcohol and other drugs on psychomotor performance and cognitive function. Alcohol and Alcoholism 26: 71–79, 1991

    PubMed  Google Scholar 

  9. Fuches V, Coper H, Romelspacher T: The effect of ethanol and haloperiodol on dopamine receptor (D) density. Neuropharmacology 26: 1231–1233, 1987

    PubMed  Google Scholar 

  10. Babu PP, Nagaraju N, Vemuri MC: Differences in the plasma membrane proteins of chronic alcoholic rat brain. Membrane Biochemistry 9: 227–237, 1990

    PubMed  Google Scholar 

  11. Haviryaji KSG, Srinivas R, Suryanarayana T, Vemuri MC: Effect of ethanol on hepatic ribosomal proteins and mRNA. Molecular and Cellular Biochemistry 115: 143–147, 1992

    PubMed  Google Scholar 

  12. Farooq M, Norton WJ: A modified procedure for isolation of astrocyte and neuron enriched fractions from rat brain. J Neurochem 31: 887–894, 1978

    PubMed  Google Scholar 

  13. Pinkett O, Anderson WB: Alterations in plasma membrane proteins associated with the proteolytic activation of adenylate cyclase. Arch Biochem Biophys 200: 261–268, 1980

    PubMed  Google Scholar 

  14. Gurd JW: Phosphorylation of post-synaptic density glycoprotein gp 180 by Ca2+/calmodulin-dependent protein kinase. J Neurochem 45: 1128–1135, 1985

    PubMed  Google Scholar 

  15. Ali SM, Bullock S, Rose SPR: Phosphorylation of synaptic proteins in chick forebrain: Changes with development and passive avoidance training. J Neurochem 50: 1579–1587, 1988

    PubMed  Google Scholar 

  16. Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685, 1970

    PubMed  Google Scholar 

  17. Cheng YE, Chen LB: Detection of phosphotyrosine-containing 34,000-dalton protein in the framework of cells transformed with Rous sarcoma virus. Proc Natl Acad Sci USA 78: 2388–2392, 1981

    PubMed  Google Scholar 

  18. Kawahara H, Cadrin M, French SW: Ethanol-induced phosphorylation of cytokeratin in cultured hepatocytes. Life Sciences 47: 859–863, 1990

    PubMed  Google Scholar 

  19. Tardivel S, Dupuis Y, Fournier P, Zelazowska E, Grabon W, Porembska Z: Differential effect of ethanol on formation of phosphorylated intermediates of the Jejunal brush border alkaline phosphatase Mr 90,000 and 65,000 subunits. Acta Biochem Pol 37: 177–180, 1990

    Google Scholar 

  20. Harrison BC, Mobley PL: Protein phosphorylation in astrocytes mediated by protein kinase C: Comparison with phosphorylation by cyclic AMP-dependent protein kinase. J Neurochem 53: 1245–1251, 1989

    PubMed  Google Scholar 

  21. Nishizuka Y: Studies and prospectives of the protein kinase C family for cellular regulation. Cancer 63: 1892–1903, 1989

    PubMed  Google Scholar 

  22. Masliah E, Yoshida K, Shimohama S, Gage FH, Saitoh T: Differential expression of protein kinase C isoenzymes in rat glial cell cultures. Brain Res 549: 106–111, 1991

    PubMed  Google Scholar 

  23. Kikkawa U, Kishimoto A, Nishizuka Y: The protein kinase C family: Heterogeneity and its implications. Annu Rev Biochem 58: 31–44, 1989

    PubMed  Google Scholar 

  24. Murphy S, McCabe N, Morrow C, Pearce B: Phorbol ester stimulates proliferation of astrocytes in primary culture. Dev Brain Res 31: 133–135, 1987

    Google Scholar 

  25. Nhamburo PT, Salafsky BP, Tabakoff B, Hoffman PL: Effects of ethanol on ouabain inhibition of mouse brain (Na+, K+) ATPase activity. Biochem Pharmacol 36: 2027–2033, 1987

    PubMed  Google Scholar 

  26. Charness ME, Querimit LA, Henteleff M: Ethanol differentially regulates G proteins in neural cells. Biochem Biophys Res Commun 155: 138–141, 1988

    PubMed  Google Scholar 

  27. Messing RO, Petersen PJ, Henrich CJ: Chronic ethanol exposure increases levels of protein kinase C Δ and ɛ and protein kinase C-mediated phosphorylation in cultured neural cells. J Biol Chem 266: 23428–23432, 1991

    PubMed  Google Scholar 

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Babu, P.P., Kumari, L.R. & Vemuri, M.C. Ethanol induced alterations in plasma membrane protein phosphorylation of neurons and astrocytes. Mol Cell Biochem 130, 41–48 (1994). https://doi.org/10.1007/BF01084266

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  • DOI: https://doi.org/10.1007/BF01084266

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