Skip to main content
SpringerLink
Log in

Tyrosinated, detyrosinated and acetylated tubulin isotypes in rat brain membranes. Their proportions in comparison with those in cytosol

  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

The heterogeneity of α-tubulin and the relative proportions of the tubulin isotypes were investigated in brain membranes of rats of 1, 25 and 180 days of age by using four anti-α-tubulin antibodies: a) the monoclonal DM1A antibody, specific for α-tubulin; b) the monoclonal 6-11B-1 antibody, specific for acetylated tubulin; c) a polyclonal antibody (Glu antibody), specific for detyrosinated tubulin; and d) a polyclonal antibody (Tyr antibody), specific for tyrosinated tubulin. We found that rat brain membranes contain the three tubulin isotypes mentioned above. The proportions of tyrosinated and detyrosinated tubulin relative to total α-tubulin were somewhat lower in membrane than in cytosol in animals of 25 and 180 days of age. At day one of development, the proportions in membrane were similar to those found in cytosol. With respect to the acetylated form, it was about 20 times higher in membrane than in cytosol at the three ages studied. The proportion of acetylated tubulin was determined in different subcellular fractions: myelin, synaptic vesicles, mitochondria, microsomes, and plasma membrane. While the amount of total tubulin differed between the different subcellular fractions, the proportion of acetylated tubulin relative to total α-tubulin was constant and similar to that found in total membranes. The proportion of acetylated tubulin was also investigated in non-neural tissues (kidney, liver and lung). Although values for cytosol were about 10-fold higher than that found in brain cytosol, no detectable values for membranes could be obtained in these organs.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Cleveland DW, Sullivan KF: Molecular biology and genetics of tubulin. Ann Rev Biochem 54: 31–365, 1985

    Google Scholar 

  2. Lewis SA, Lee MGS, Cowan NJ: Five mouse tubulin isotypes and their regulated expression during development. J Cell Biol 101: 852–861, 1985

    Google Scholar 

  3. Gard DL, Kirschner MW: A polymer-dependent increase in phosphorylation of β-tubulin accompanies differentiation of a mouse neuroblastoma cell line. J Cell Biol 100: 765–774, 1985

    Google Scholar 

  4. Matten WT, Aubry M, West J, Maness P: Tubulin is phosphorylated at tyrosine by pp60c-src in nerve growth cone membranes. J Cell Biol 111: 1959–1970, 1990

    Google Scholar 

  5. Barra HS, Arce CA, Argaraña CE: Posttranslational tyrosination/detyrosination of tubulin. Mol Neurobiol 2: 133–153, 1988

    Google Scholar 

  6. L'Hernault SW, Rosenbaum JL: Chlamydomonas α-tubulin is posttranslationally modified by acetylation on the ɛ-amino group of a lysine. Biochemistry 24: 473–478, 1985

    Google Scholar 

  7. LeDizet M, Piperno G: Identification of an acetylation site of Chlamydomonas α-tubulin. Proc Natl Acad Sci USA 84: 5720–5724, 1987

    Google Scholar 

  8. Eddé B, Rossier J, Le Caer JP, Desbruyeres E, Gros F, Denoulet P: Posttranslational glutamylation of α-tubulin. Science 247: 83–85, 1990

    Google Scholar 

  9. Joshi HC, Cleveland DW: Diversity among tubulin subunits: toward what functional end? Cell Motil Cytoskel 16: 159–163, 1990

    Google Scholar 

  10. Walters BB, Matus AI: Tubulin in postsynaptic functional lattice. Nature 257: 496–498, 1975

    Google Scholar 

  11. Gozes I, Littauer U: The alfa subunit of tubulin is preferentially associated with brain presynaptic membrane. FEBS Lett 99: 86–90, 1979

    Google Scholar 

  12. Strocchi P, Brown BA, Young JD, Bonventre JA, Gilbert JM: The characterization of tubulin in CNS membrane fractions. J Neurochem 37: 1295–1307, 1981

    Google Scholar 

  13. Bernier-Valentin F, Aunis D, Rousset B: Evidence for tubulin binding sites on cellular membranes: plasma membranes, mitochondrial membranes, and secretory granule membranes. J Cell Biol 97: 209–216, 1983

    Google Scholar 

  14. de Néchaud B, Wolff A, Jeantet C, Bourre JM: Characterization of tubulin in mouse brain myelin. J Neurochem 41: 1538–1544, 1983

    Google Scholar 

  15. Hargreaves AJ, Avila J: Localization and characterization of tubulin-like proteins associated with brain mitochondria: The presence of a membrane-specific isoform. J Neurochem 45: 490–496, 1985

    Google Scholar 

  16. Stephens RE: Evidence for a tubulin-containing lipid-protein structural complex in ciliary membranes. J Cell Biol 100: 1082–1090, 1985

    Google Scholar 

  17. Regula CS, Sager PR, Berlin RD: Membrane tubulin. In: D Soifer (ed) Dynamic Aspects of Microtubule Biology. Ann NY Acad Sci 466: 832–842, 1986

  18. Lewis SA, Cowan N: Complex regulation and functional versatility of mammalian α- and β-tubulin isotypes during the differentiation of testis and muscle cells. J Cell Biol 106: 2023–2033, 1988

    Google Scholar 

  19. Sullivan K: Structure and utilization of tubulin isotypes. Ann Rev Cell Biol 4: 687–716, 1988

    Google Scholar 

  20. Gundersen GG, Kalnoski MH, Bulinski JC: Distinct populations of microtubules: Tyrosinated and nontyrosinated alpha tubulin are distributed differently in vivo. Cell 38: 779–789, 1984

    Google Scholar 

  21. Bordier C: Phase separation of integral membrane proteins in Triton X-114 solution. J Biol Chem 256: 1604–1607, 1981

    Google Scholar 

  22. Sloboda RD, Rosenbaum JL: Purification and assay of microtubule-associated proteins (MAPS). Methods Enzymol 85: 409–416, 1982

    Google Scholar 

  23. Beltramo DM, Arce CA, Barra HS: Tyrosination-detyrosination of tubulin and microtubules during the development of chick erythrocytes. Mol Cell Biochem 89: 47–56, 1989

    Google Scholar 

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

    PubMed  Google Scholar 

  25. Towbin H, Staehelin T, Gordon J: Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications. Proc Nat Acad Sci 76: 4350–4354, 1979

    Google Scholar 

  26. Greenwood FC, Hunter WM, Glover JS: The preparation of 131I-labeled human growth hormone of high specific radioactivity. Biochem J 89: 114–123, 1963

    Google Scholar 

  27. Bradford MM: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248–254, 1976

    Article  CAS  PubMed  Google Scholar 

  28. Whittaker VP, Michaelson IA, Kirkland RJA: The separation of synaptic vesicles from nerve-ending particles (synaptosomes). Biochem J 90: 293–303, 1964

    Google Scholar 

  29. Barra HS, Arce CA, Caputto R: Total tubulin and its aminoacylated and non-aminoacylated forms during development of rat brain. Eur J Biochem 109: 439–446, 1980

    Google Scholar 

  30. Cambray-Deakin MA, Burgoyne RD: Posttranslational modifications of α-tubulin: Acetylated and detyrosinated forms in axons of rat cerebellum. J Cell Biol 104: 1569–1574, 1987

    Google Scholar 

  31. Blose SH, Meltzer DI, Feramisco JR: 10-nm filaments are induced to collapse in living cells microinjected with monoclonal and polyclonal antibodies against tubulin. J Cell Biol 98: 847–858, 1984

    Google Scholar 

  32. Piperno G, Fuller MT: Monoclonal antibodies specific for an acetylated form of α-tubulin recognize the antigen in cilia and flagella from a variety of organisms. J Cell Biol 101: 2085–2094, 1985

    Google Scholar 

  33. Maruta H, Greer K, Rosenbaum JL: The acetylation of alphatubulin and its relationship to the assembly and disassembly of microtubules. J Cell Biol 103: 571–579, 1986

    Google Scholar 

  34. Black MM, Keyser P: Acetylation of α-tubulin in cultured neurons (Abstr). J Cell Biol 103: 128, 1986

    Google Scholar 

  35. Gupta RS: Mitochondria, molecular chaperone proteins and the in vivo assembly of microtubules. Trends Biochem Sci 15: 415–418, 1990

    Google Scholar 

  36. Stephens RE: Membrane tubulin. Biol Cell 57: 95–109, 1986

    Google Scholar 

  37. Ravindra R, Aronstam RS: Influence of anti-tubulin antibodies on muscarinic receptor modulation of G protein GTPase activity in rat striatum. Biochem Pharmacol 40: 457–463, 1990

    Google Scholar 

  38. Houdebine LM: The possible involvement of tubulin in transduction of the prolactin signal. Reprod Nutr Dev 30: 431–438, 1990

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Centro de Investigaciones en Química Biológica de Córdoba, Facultad de Ciencias Químicas, Universidad Nacional de Córdoba, 5016, Casilla de Correo 61, Córdoba, Argentina

    Dante M. Beltramo, Alejandra del C. Alonso & Héctor S. Barra

Authors
  1. Dante M. Beltramo
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Alejandra del C. Alonso
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Héctor S. Barra
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Beltramo, D.M., Alonso, A.d.C. & Barra, H.S. Tyrosinated, detyrosinated and acetylated tubulin isotypes in rat brain membranes. Their proportions in comparison with those in cytosol. Mol Cell Biochem 112, 173–180 (1992). https://doi.org/10.1007/BF00227574

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00227574

Key words

Search

Navigation