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. 1981 Sep 1;90(3):755–760. doi: 10.1083/jcb.90.3.755

Preferential phosphorylation of the 150,000 molecular weight component of neurofilaments by a cyclic AMP-dependent, microtubule-associated protein kinase

PMCID: PMC2111909  PMID: 6270160

Abstract

Highly purified preparations of bovine brain and rabbit nerve root neurofilaments were found to be lacking in protein kinase activity when either histone FIIA or the neurofilaments themselves were used as acceptors. There was no augmentation of activity in the presence of cyclic AMP. Addition of microtubule proteins prepared by cycles of assembly and disassembly resulted in phosphorylation of histone, phosphorylation of tubulin and the microtubule-associated proteins, and phosphorylation of neurofilament subunits. The phosphorylation of neurofilaments was predominantly in the 150,000-dalton species and was completely cyclic AMP dependent.

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Selected References

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  1. Berkowitz S. A., Katagiri J., Binder H. K., Williams R. C., Jr Separation and characterization of microtubule proteins from calf brain. Biochemistry. 1977 Dec 13;16(25):5610–5617. doi: 10.1021/bi00644a035. [DOI] [PubMed] [Google Scholar]
  2. Cleveland D. W., Hwo S. Y., Kirschner M. W. Physical and chemical properties of purified tau factor and the role of tau in microtubule assembly. J Mol Biol. 1977 Oct 25;116(2):227–247. doi: 10.1016/0022-2836(77)90214-5. [DOI] [PubMed] [Google Scholar]
  3. Cooke P. A filamentous cytoskeleton in vertebrate smooth muscle fibers. J Cell Biol. 1976 Mar;68(3):539–556. doi: 10.1083/jcb.68.3.539. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Eng L. F., Vanderhaeghen J. J., Bignami A., Gerstl B. An acidic protein isolated from fibrous astrocytes. Brain Res. 1971 May 7;28(2):351–354. doi: 10.1016/0006-8993(71)90668-8. [DOI] [PubMed] [Google Scholar]
  5. Franke W. W., Schmid E., Winter S., Osborn M., Weber K. Widespread occurrence of intermediate-sized filaments of the vimentin-type in cultured cells from diverse vertebrates. Exp Cell Res. 1979 Oct 1;123(1):25–46. doi: 10.1016/0014-4827(79)90418-x. [DOI] [PubMed] [Google Scholar]
  6. Goodman D. B., Rasmussen H., DiBella F., Guthrow C. E., Jr Cyclic adenosine 3':5'-monophosphate-stimulated phosphorylation of isolated neurotubule subunits. Proc Natl Acad Sci U S A. 1970 Oct;67(2):652–659. doi: 10.1073/pnas.67.2.652. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Izant J. G., McIntosh J. R. Microtubule-associated proteins: a monoclonal antibody to MAP2 binds to differentiated neurons. Proc Natl Acad Sci U S A. 1980 Aug;77(8):4741–4745. doi: 10.1073/pnas.77.8.4741. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Karr T. L., White H. D., Purich D. L. Characterization of brain microtubule proteins prepared by selective removal of mitochondrial and synaptosomal components. J Biol Chem. 1979 Jul 10;254(13):6107–6111. [PubMed] [Google Scholar]
  9. 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]
  10. Lasek R. J. The dynamic ordering of neuronal cytoskeletons. Neurosci Res Program Bull. 1981 Feb;19(1):7–32. [PubMed] [Google Scholar]
  11. Lazarides E. Intermediate filaments as mechanical integrators of cellular space. Nature. 1980 Jan 17;283(5744):249–256. doi: 10.1038/283249a0. [DOI] [PubMed] [Google Scholar]
  12. Liem R. K., Yen S. H., Salomon G. D., Shelanski M. L. Intermediate filaments in nervous tissues. J Cell Biol. 1978 Dec;79(3):637–645. doi: 10.1083/jcb.79.3.637. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Peters A., Vaughn J. E. Microtubules and filaments in the axons and astrocytes of early postnatal rat optic nerves. J Cell Biol. 1967 Jan;32(1):113–119. doi: 10.1083/jcb.32.1.113. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Rappaport L., Leterrier J. F., Virion A., Nunez J., Osty J. Phosphorylation of microtubule-associated proteins. Eur J Biochem. 1976 Mar 1;62(3):539–549. doi: 10.1111/j.1432-1033.1976.tb10188.x. [DOI] [PubMed] [Google Scholar]
  15. Runge M. S., Schlaepfer W. W., Williams R. C., Jr Isolation and characterization of neurofilaments from mammalian brain. Biochemistry. 1981 Jan 6;20(1):170–175. doi: 10.1021/bi00504a028. [DOI] [PubMed] [Google Scholar]
  16. Runge M. S., el-Maghrabi M. R., Claus T. H., Pilkis S. J., Williams R. C., Jr A MAP-2-stimulated protein kinase activity associated with neurofilaments. Biochemistry. 1981 Jan 6;20(1):175–180. doi: 10.1021/bi00504a029. [DOI] [PubMed] [Google Scholar]
  17. Schlaepfer W. W., Freeman L. A. Neurofilament proteins of rat peripheral nerve and spinal cord. J Cell Biol. 1978 Sep;78(3):653–662. doi: 10.1083/jcb.78.3.653. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Shecket G., Lasek R. J. Preparation of neurofilament protein from guinea pig peripheral nerve and spinal cord. J Neurochem. 1980 Dec;35(6):1335–1344. doi: 10.1111/j.1471-4159.1980.tb09007.x. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Shelanski M. L., Leterrier J. F., Liem R. K. Evidence for interactions between neurofilaments and microtubules. Neurosci Res Program Bull. 1981 Feb;19(1):32–43. [PubMed] [Google Scholar]
  21. Shelanski M. L., Liem R. K. Neurofilaments. J Neurochem. 1979 Jul;33(1):5–13. doi: 10.1111/j.1471-4159.1979.tb11699.x. [DOI] [PubMed] [Google Scholar]
  22. Sloboda R. D., Rudolph S. A., Rosenbaum J. L., Greengard P. Cyclic AMP-dependent endogenous phosphorylation of a microtubule-associated protein. Proc Natl Acad Sci U S A. 1975 Jan;72(1):177–181. doi: 10.1073/pnas.72.1.177. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Sun T. T., Green H. Keratin filaments of cultured human epidermal cells. Formation of intermolecular disulfide bonds during terminal differentiation. J Biol Chem. 1978 Mar 25;253(6):2053–2060. [PubMed] [Google Scholar]
  24. Tsukita S., Ishikawa H. The movement of membranous organelles in axons. Electron microscopic identification of anterogradely and retrogradely transported organelles. J Cell Biol. 1980 Mar;84(3):513–530. doi: 10.1083/jcb.84.3.513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. 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]
  26. Walsh D. A., Ashby C. D., Gonzalez C., Calkins D., Fischer E. H. Krebs EG: Purification and characterization of a protein inhibitor of adenosine 3',5'-monophosphate-dependent protein kinases. J Biol Chem. 1971 Apr 10;246(7):1977–1985. [PubMed] [Google Scholar]
  27. Wisniewski H., Shelanski M. L., Terry R. D. Effects of mitotic spindle inhibitors on neurotubules and neurofilaments in anterior horn cells. J Cell Biol. 1968 Jul;38(1):224–229. doi: 10.1083/jcb.38.1.224. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Yen S. H., Liem R. K., Jenq L. T., Shelanski M. L. Rapid purification of intact tonofilaments from newborn rats. Comparison with glial filaments and neurofilaments. Exp Cell Res. 1980 Oct;129(2):313–320. doi: 10.1016/0014-4827(80)90498-x. [DOI] [PubMed] [Google Scholar]

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