Skip to main content
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1986 Mar 1;102(3):1060–1066. doi: 10.1083/jcb.102.3.1060

Identification of a 34-kD polypeptide as a light chain of microtubule- associated protein-1 (MAP-1) and its association with a MAP-1 peptide that binds to microtubules

PMCID: PMC2114103  PMID: 3512577

Abstract

We examined the association of a 34-kD light chain component to the heavy chains of MAP-1 using a monoclonal antibody that specifically binds the 34-kD component and labels neuronal microtubules in a specific and saturable manner. Immunoprecipitation of MAP-1 heavy chains together with the 34-kD component by the antibody indicates that the 34-kD polypeptide forms a complex with MAP-1 heavy chains. Both major isoforms of MAP-1 heavy chains (MAP-1A and MAP-1B) were found in the immunoprecipitate. Digestion of MAP-1 with alpha-chymotrypsin and analysis of the chymotryptic peptides reveals a 120-kD fragment of the MAP-1 heavy chain that binds to microtubules and is precipitable with the 34-kD light chain antibody, suggesting that the 34-kD light chain also binds to this domain of the molecule. Since microtubules that contain the 120-kD fragment lack the long lateral projections characteristic of microtubules with intact MAP-1, the 34-kD light chains may be localized at or near the microtubule surface.

Full Text

The Full Text of this article is available as a PDF (891.5 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bloom G. S., Luca F. C., Vallee R. B. Widespread cellular distribution of MAP-1A (microtubule-associated protein 1A) in the mitotic spindle and on interphase microtubules. J Cell Biol. 1984 Jan;98(1):331–340. doi: 10.1083/jcb.98.1.331. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bloom G. S., Schoenfeld T. A., Vallee R. B. Widespread distribution of the major polypeptide component of MAP 1 (microtubule-associated protein 1) in the nervous system. J Cell Biol. 1984 Jan;98(1):320–330. doi: 10.1083/jcb.98.1.320. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. 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]
  4. Kotani S., Murofushi H., Nishida E., Sakai H. 33K protein--an inhibitory factor of tubulin polymerization in porcine brain. J Biochem. 1984 Oct;96(4):959–969. doi: 10.1093/oxfordjournals.jbchem.a134955. [DOI] [PubMed] [Google Scholar]
  5. Kumagai H., Sakai H. A porcine brain protein (35 K protein) which bundles microtubules and its identification as glyceraldehyde 3-phosphate dehydrogenase. J Biochem. 1983 May;93(5):1259–1269. doi: 10.1093/oxfordjournals.jbchem.a134260. [DOI] [PubMed] [Google Scholar]
  6. Kuznetsov S. A., Rodionov V. I., Bershadsky A. D., Gelfand V. I., Rosenblat V. A. High molecular weight protein MAP 2 promoting microtubule assembly in vitro is associated with microtubules in cells. Cell Biol Int Rep. 1980 Nov;4(11):1017–1024. doi: 10.1016/0309-1651(80)90174-5. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Kuznetsov S. A., Rodionov V. I., Gelfand V. I., Rosenblat V. A. Microtubule-associated protein MAP1 promotes microtubule assembly in vitro. FEBS Lett. 1981 Dec 7;135(2):241–244. doi: 10.1016/0014-5793(81)80791-0. [DOI] [PubMed] [Google Scholar]
  9. Kuznetsov S. A., Rodionov V. I., Gelfand V. I., Rosenblat V. A. Purification of high-Mr microtubule proteins MAP1 and MAP2. FEBS Lett. 1981 Dec 7;135(2):237–240. doi: 10.1016/0014-5793(81)80790-9. [DOI] [PubMed] [Google Scholar]
  10. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  11. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  12. Matus A., Bernhardt R., Hugh-Jones T. High molecular weight microtubule-associated proteins are preferentially associated with dendritic microtubules in brain. Proc Natl Acad Sci U S A. 1981 May;78(5):3010–3014. doi: 10.1073/pnas.78.5.3010. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. REYNOLDS E. S. The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J Cell Biol. 1963 Apr;17:208–212. doi: 10.1083/jcb.17.1.208. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Rodionov V. I., Nadezhdina E. S., Leonova E. V., Vaisberg E. A., Kuznetsov S. A., Gelfand V. I. Identification of a 100 kD protein associated with microtubules, intermediate filaments and coated vesicles in cultured cells. Exp Cell Res. 1985 Aug;159(2):377–387. doi: 10.1016/s0014-4827(85)80011-2. [DOI] [PubMed] [Google Scholar]
  15. Shelanski M. L., Gaskin F., Cantor C. R. Microtubule assembly in the absence of added nucleotides. Proc Natl Acad Sci U S A. 1973 Mar;70(3):765–768. doi: 10.1073/pnas.70.3.765. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Sloboda R. D., Dickersin K. Structure and composition of the cytoskeleton of nucleated erythrocytes I. The presence of microtubule-associated protein 2 in the marginal band. J Cell Biol. 1980 Oct;87(1):170–179. doi: 10.1083/jcb.87.1.170. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Stearns M. E. Cytomatrix in chromatophores. J Cell Biol. 1984 Jul;99(1 Pt 2):144s–151s. doi: 10.1083/jcb.99.1.144s. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Vallee R. B., Bloom G. S., Theurkauf W. E. Microtubule-associated proteins: subunits of the cytomatrix. J Cell Biol. 1984 Jul;99(1 Pt 2):38s–44s. doi: 10.1083/jcb.99.1.38s. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Vallee R. B., Davis S. E. Low molecular weight microtubule-associated proteins are light chains of microtubule-associated protein 1 (MAP 1). Proc Natl Acad Sci U S A. 1983 Mar;80(5):1342–1346. doi: 10.1073/pnas.80.5.1342. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Vallee R. Structure and phosphorylation of microtubule-associated protein 2 (MAP 2). Proc Natl Acad Sci U S A. 1980 Jun;77(6):3206–3210. doi: 10.1073/pnas.77.6.3206. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Weatherbee J. A., Sherline P., Mascardo R. N., Izant J. G., Luftig R. B., Weihing R. R. Microtubule-associated proteins of HeLa cells: heat stability of the 200,000 mol wt HeLa MAPs and detection of the presence of MAP-2 in HeLa cell extracts and cycled microtubules. J Cell Biol. 1982 Jan;92(1):155–163. doi: 10.1083/jcb.92.1.155. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]
  24. 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]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES