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PMID: 3091608 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

Identification of a MAP 2-like ATP-binding protein associated with axoplasmic vesicles that translocate on isolated microtubules.

The Journal of cell biology ·Vol. 103 ·No. 3 ·1986-09-00 ·Pages 947-56

Gilbert SP, Sloboda RD

Abstract

Axoplasmic vesicles were purified and observed to translocate on isolated microtubules in an ATP-dependent, trypsin-sensitive manner, implying that ATP-binding polypeptides essential for force generation were present on the vesicle surface. To identify these proteins [alpha 32P]8-azidoadenosine 5'-triphosphate ([alpha 32P]8-N3ATP), a photoaffinity analogue of ATP, was used. The results presented here identify and characterize a vesicle-associated polypeptide having a relative molecular mass of 292 kD that bound [alpha 32P]8-N3ATP. The incorporation of label is ultraviolet light-dependent and ATP-sensitive. Moreover, the 292-kD polypeptide could be isolated in association with vesicles or microtubules, depending on the conditions used, and the data indicate that the 292-kD polypeptide is similar to mammalian brain microtubule-associated protein 2 (MAP 2) for the following reasons: The 292-kD polypeptide isolated from either squid axoplasm or optic lobe cross-reacts with antiserum to porcine brain MAP 2. Furthermore, it purifies with taxol-stabilized microtubules and is released with salt. Based on these characteristics, the 292-kD polypeptide is distinct from the known force-generating molecules myosin and flagellar dynein, as well as the 110-130-kD kinesin-like polypeptides that have recently been described (Brady, S. T., 1985, Nature (Lond.), 317:73-75; Vale, R. D., T. S. Reese, and M. P. Sheetz, 1985b, Cell, 42:39-50; Scholey, J. M., M. E. Porter, P. M. Grissom, and J. R. McIntosh, 1985, Nature (Lond.), 318:483-486). Because the 292-kD polypeptide binds ATP and is associated with vesicles that translocate on purified MAP-free microtubules in an ATP-dependent fashion, it is therefore believed to be involved in vesicle-microtubule interactions that promote organelle motility.

MeSH Terms
Adenosine Triphosphate/metabolism Affinity Labels Animals Decapodiformes Immunoelectrophoresis Kinesins Microtubule-Associated Proteins/isolation & purification Microtubules/analysis Movement Nerve Tissue Proteins/analysis Optic Lobe, Nonmammalian/analysis Sea Urchins
Chemicals
Affinity Labels Microtubule-Associated Proteins Nerve Tissue Proteins Adenosine Triphosphate Kinesins
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Gilbert S P
Sloboda R D
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.103.3.947
Published
1986-09-00
Pages
947-56
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2114312
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.103.3.947 PMC Full Text View journal details
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