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PMID: 4055896 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Analysis of the microtubule-binding domain of MAP-2.

The Journal of cell biology ·Vol. 101 ·No. 5 Pt 1 ·1985-11-00 ·Pages 1782-9

Gottlieb RA, Murphy DB

Abstract

We examined the microtubule-binding domain of the microtubule-associated protein (MAP), MAP-2, using rabbit antibodies that specifically bind to the microtubule-binding region ("stub") and the projection portion ("arm") of MAP-2. We found that (a) microtubules decorated with arm antibody look similar to those labeled with whole unfractionated MAP antibody, though microtubules are not labeled with stub antibody; (b) incubation of depolymerized microtubule protein with stub antibody prior to assembly partially inhibits the rate of microtubule elongation, presumably because MAPs that are complexed with antibody cannot bind to microtubules and stabilize elongating polymers; (c) the rate of appearance and amounts of 36- and 40-kD microtubule-binding peptides produced by digestion with chymotrypsin are distinct for MAPs associated with microtubules vs. MAPs free in solution. The enhanced stability of the 40-kD peptide when associated with microtubules suggests that this domain of the protein is closely associated with, or partially buried in, the microtubule surface; (d) MAP-2 is a slender, elongate molecule as determined by unidirectional platinum shadowing (90 +/- 30 nm), which is in approximate agreement with previous observations. Stub antibody labels MAP-2 in the terminal one-quarter of the extended protein, indicating an intrinsic asymmetry in the molecule.

MeSH Terms
Animals Antibodies Binding Sites Brain/metabolism Chromatography, Affinity Chymotrypsin/metabolism Microscopy, Electron Microtubule-Associated Proteins/isolation & purification,metabolism Microtubules/metabolism,ultrastructure Peptide Fragments/analysis Protein Binding Swine
Chemicals
Antibodies Microtubule-Associated Proteins Peptide Fragments Chymotrypsin
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Gottlieb R A
Murphy D B
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29 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.101.5.1782
Published
1985-11-00
Pages
1782-9
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2113981
Subset
IM
Grants
NCI NIH HHS · CA00645 · United States
NIGMS NIH HHS · GM26155 · United States
External Links
PubMed source DOI: 10.1083/jcb.101.5.1782 PMC Full Text View journal details
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