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. 1988 Jul;7(7):1957–1963. doi: 10.1002/j.1460-2075.1988.tb03033.x

Differential interaction of synthetic peptides from the carboxyl-terminal regulatory domain of tubulin with microtubule-associated proteins.

R B Maccioni 1, C I Rivas 1, J C Vera 1
PMCID: PMC454467  PMID: 3416830

Abstract

In previous studies we have demonstrated that a 4-kd tubulin fragment, including amino acid residues from Phe418 to Glu450 in alpha-subunit and Phe408-Ala445 of the beta-sequence, plays a major role in controlling tubulin interactions leading to microtubule assembly. The 4-kd carboxyl-terminal domain also constitutes an essential domain for the interaction of microtubule-associated proteins (MAPs). Removal of the 4-kd fragment facilitates tubulin self-association and renders the assembly MAP-independent. In order to define the substructure of the tubulin domain for MAP interaction, we have examined the binding of 3H-acetylated C-terminal peptides to MAP-2 and tau. Two synthetic peptides from the low-homology region within the 4-kd domain alpha (430-441) and beta (422-434) and the peptide, alpha (401-410) of the high-homology region adjacent to the 4-kd domain, were analyzed with respect to MAP interaction. The binding data showed a relatively strong interaction of MAP-2 with the beta (422-434) peptide and a weaker interaction of both MAPs components with alpha (430-441) tubulin peptide as analyzed by Airfuge ultracentrifugation and zone filtration chromatography. The homologous alpha (401-410) peptide did not bind to either MAP-2 or tau. Equilibrium dialysis experiments showed a co-operative binding of beta (422-434) peptide to multiple sites in tau. The alpha (430-441) peptide exhibited a stronger interaction for tau as compared with MAP-2.(ABSTRACT TRUNCATED AT 250 WORDS)

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Selected References

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  1. Bond J. F., Fridovich-Keil J. L., Pillus L., Mulligan R. C., Solomon F. A chicken-yeast chimeric beta-tubulin protein is incorporated into mouse microtubules in vivo. Cell. 1986 Feb 14;44(3):461–468. doi: 10.1016/0092-8674(86)90467-8. [DOI] [PubMed] [Google Scholar]
  2. Breitling F., Little M. Carboxy-terminal regions on the surface of tubulin and microtubules. Epitope locations of YOL1/34, DM1A and DM1B. J Mol Biol. 1986 May 20;189(2):367–370. doi: 10.1016/0022-2836(86)90517-6. [DOI] [PubMed] [Google Scholar]
  3. Carlier M. F., Simon C., Pantaloni D. Polymorphism of tubulin oligomers in the presence of microtubule-associated proteins. Implications in microtubule assembly. Biochemistry. 1984 Mar 27;23(7):1582–1590. doi: 10.1021/bi00302a037. [DOI] [PubMed] [Google Scholar]
  4. Cleveland D. W. The multitubulin hypothesis revisited: what have we learned? J Cell Biol. 1987 Mar;104(3):381–383. doi: 10.1083/jcb.104.3.381. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Fridovich-Keil J. L., Bond J. F., Solomon F. Domains of beta-tubulin essential for conserved functions in vivo. Mol Cell Biol. 1987 Oct;7(10):3792–3798. doi: 10.1128/mcb.7.10.3792. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Furtner R., Wiche G. Binding specificities of purified porcine brain alpha- and beta-tubulin subunits and of microtubule-associated proteins 1 and 2 examined by electron microscopy and solid-phase binding assays. Eur J Cell Biol. 1987 Dec;45(1):1–8. [PubMed] [Google Scholar]
  7. Gaskin F., Cantor C. R., Shelanski M. L. Turbidimetric studies of the in vitro assembly and disassembly of porcine neurotubules. J Mol Biol. 1974 Nov 15;89(4):737–755. doi: 10.1016/0022-2836(74)90048-5. [DOI] [PubMed] [Google Scholar]
  8. Havercroft J. C., Cleveland D. W. Programmed expression of beta-tubulin genes during development and differentiation of the chicken. J Cell Biol. 1984 Dec;99(6):1927–1935. doi: 10.1083/jcb.99.6.1927. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hernández M. A., Avila J., Andreu J. M. Physicochemical characterization of the heat-stable microtubule-associated protein MAP2. Eur J Biochem. 1986 Jan 2;154(1):41–48. doi: 10.1111/j.1432-1033.1986.tb09356.x. [DOI] [PubMed] [Google Scholar]
  10. Herzog W., Weber K. Fractionation of brain microtubule-associated proteins. Isolation of two different proteins which stimulate tubulin polymerization in vitro. Eur J Biochem. 1978 Dec 1;92(1):1–8. doi: 10.1111/j.1432-1033.1978.tb12716.x. [DOI] [PubMed] [Google Scholar]
  11. Howlett G. J., Yeh E., Schachman H. K. Protein-ligand binding studies with a table-top, air-driven high-speed centrifuge. Arch Biochem Biophys. 1978 Oct;190(2):808–819. doi: 10.1016/0003-9861(78)90341-7. [DOI] [PubMed] [Google Scholar]
  12. Kuznetsov S. A., Rodionov V. I., Gelfand V. I., Rosenblat V. A. MAP2 competes with MAP1 for binding to microtubules. Biochem Biophys Res Commun. 1984 Feb 29;119(1):173–178. doi: 10.1016/0006-291x(84)91635-8. [DOI] [PubMed] [Google Scholar]
  13. Lee G., Cowan N., Kirschner M. The primary structure and heterogeneity of tau protein from mouse brain. Science. 1988 Jan 15;239(4837):285–288. doi: 10.1126/science.3122323. [DOI] [PubMed] [Google Scholar]
  14. Lewis S. A., Gu W., Cowan N. J. Free intermingling of mammalian beta-tubulin isotypes among functionally distinct microtubules. Cell. 1987 May 22;49(4):539–548. doi: 10.1016/0092-8674(87)90456-9. [DOI] [PubMed] [Google Scholar]
  15. Littauer U. Z., Giveon D., Thierauf M., Ginzburg I., Ponstingl H. Common and distinct tubulin binding sites for microtubule-associated proteins. Proc Natl Acad Sci U S A. 1986 Oct;83(19):7162–7166. doi: 10.1073/pnas.83.19.7162. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lopata M. A., Cleveland D. W. In vivo microtubules are copolymers of available beta-tubulin isotypes: localization of each of six vertebrate beta-tubulin isotypes using polyclonal antibodies elicited by synthetic peptide antigens. J Cell Biol. 1987 Oct;105(4):1707–1720. doi: 10.1083/jcb.105.4.1707. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Maccioni R. B., Cann J. R., Stewart J. M. Interaction of substance P with tubulin. Eur J Biochem. 1986 Jan 15;154(2):427–435. doi: 10.1111/j.1432-1033.1986.tb09415.x. [DOI] [PubMed] [Google Scholar]
  18. Maccioni R. B., Serrano L., Avila J., Cann J. R. Characterization and structural aspects of the enhanced assembly of tubulin after removal of its carboxyl-terminal domain. Eur J Biochem. 1986 Apr 15;156(2):375–381. doi: 10.1111/j.1432-1033.1986.tb09593.x. [DOI] [PubMed] [Google Scholar]
  19. Sackett D. L., Wolff J. Proteolysis of tubulin and the substructure of the tubulin dimer. J Biol Chem. 1986 Jul 5;261(19):9070–9076. [PubMed] [Google Scholar]
  20. Serrano L., Avila J., Maccioni R. B. Controlled proteolysis of tubulin by subtilisin: localization of the site for MAP2 interaction. Biochemistry. 1984 Sep 25;23(20):4675–4681. doi: 10.1021/bi00315a024. [DOI] [PubMed] [Google Scholar]
  21. Serrano L., de la Torre J., Maccioni R. B., Avila J. Involvement of the carboxyl-terminal domain of tubulin in the regulation of its assembly. Proc Natl Acad Sci U S A. 1984 Oct;81(19):5989–5993. doi: 10.1073/pnas.81.19.5989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Wehland J., Willingham M. C., Sandoval I. V. A rat monoclonal antibody reacting specifically with the tyrosylated form of alpha-tubulin. I. Biochemical characterization, effects on microtubule polymerization in vitro, and microtubule polymerization and organization in vivo. J Cell Biol. 1983 Nov;97(5 Pt 1):1467–1475. doi: 10.1083/jcb.97.5.1467. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Weingarten M. D., Lockwood A. H., Hwo S. Y., Kirschner M. W. A protein factor essential for microtubule assembly. Proc Natl Acad Sci U S A. 1975 May;72(5):1858–1862. doi: 10.1073/pnas.72.5.1858. [DOI] [PMC free article] [PubMed] [Google Scholar]

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