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PMID: 6270156 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

A protein kinase bound to the projection portion of MAP 2 (microtubule-associated protein 2).

The Journal of cell biology ·Vol. 90 ·No. 3 ·1981-09-00 ·Pages 568-76

Vallee RB, DiBartolomeis MJ, Theurkauf WE

Abstract

In previous work we have demonstrated that the microtubule-associated protein 2 (MAP 2) molecule consists of two structural parts. One part of the molecule, referred to as the assembly-promoting domain, binds to the microtubule surface and is responsible for promoting microtubule assembly; the other represents a filamentous projection observed on the microtubule surface that may be involved in the interaction of microtubules with other cellular structures. MAP 2 is known to be specifically phosphorylated as the result of a protein kinase activity that is present in microtubule preparations. We have now found that the activity copurifies with the projection portion of MAP 2 itself. Kinase activity coeluted with MAP 2 when microtubule protein was subjected to either gel- filtration chromatography on bio-gel A-15m or ion-exchange chromatography on DEAE- Sephadex. The activity was released from microtubules by mild digestion with chymotrypsin in parallel with the removal by the protease of the MAP 2 projections from the microtubule surface. The association of the activity with the projection was demonstrated directly by gel filtration chromatography of the projections on bio-gel A-15m. Three protein species (M(r) = 39,000, 55,000, and 70,000) cofractionated with MAP 2, and two of these (M(r) = 39,000 and 55,000) may represent the subunits of an associated cyclic AMP- dependent protein kinase. The projection-associated activity was stimulated 10-fold by cyclic AMP and was inhibited more than 95 percent by the cyclic AMP-dependent protein kinase inhibitor from rabbit skeletal muscle. It appeared to represent the only significant activity associated with microtubules, almost no activity being found with tubulin, other MAPs, or the assembly-promoting domain of MAP 2, and was estimated to account for 7-22 percent of the total brain cytosolic protein kinase activity. The location of the kinase on the projection is consistent with a role in regulating the function of the projection, though other roles for the enzyme are also possible.

MeSH Terms
Animals Cattle Cyclic AMP/pharmacology Microtubule-Associated Proteins Microtubules/enzymology Phosphoprotein Phosphatases/metabolism Protein Kinases/isolation & purification,metabolism Proteins/isolation & purification,metabolism
Chemicals
Microtubule-Associated Proteins Proteins Cyclic AMP Protein Kinases Phosphoprotein Phosphatases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Vallee R B
DiBartolomeis M J
Theurkauf W E
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47 references, click to expand
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.90.3.568
Published
1981-09-00
Pages
568-76
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2111905
Subset
IM
Grants
NIGMS NIH HHS · GM 26701 · United States
External Links
PubMed source DOI: 10.1083/jcb.90.3.568 PMC Full Text View journal details
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