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PMID: 6237113 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Cytoplasmic dynein-like ATPase cross-links microtubules in an ATP-sensitive manner.

The Journal of cell biology ·Vol. 99 ·No. 4 Pt 1 ·1984-10-00 ·Pages 1251-8

Hollenbeck PJ, Suprynowicz F, Cande WZ

Abstract

We have prepared dynein-like ATPase from the eggs of the sea urchin Strongylocentrotus purpuratus using differential centrifugation and column chromatography. This ATPase preparation is inhibited by vanadate and erythro-9-(3-[2-hydroxynonyl]) adenine (EHNA) at concentrations similar to those that inhibit reactivated flagellar beating and spindle elongation in lysed cell models. Using microtubule affinity and ATP-induced release, we can purify this ATPase activity to a composition on SDS PAGE of four peptides ranging in molecular weight from 180,000-300,000. When viewed in darkfield optics, this affinity-purified ATPase caused extensive parallel bundling of microtubule-associated protein-free microtubules. These bundles were dispersed by 1 mM ATP but not by ATP gamma S or AMP-5'-adenylimidodiphosphate. The reformation of microtubule bundles after dispersal by ATP required ATP hydrolysis; bundles did not reform in the presence of 10 microM vanadate. Negative stain electron microscopy of these bundled microtubules revealed that they are arranged in parallel networks with extensive close lateral association.

MeSH Terms
Adenosine Triphosphatases/isolation & purification,metabolism Adenosine Triphosphate/metabolism Animals Cytoplasm/enzymology Dyneins Female Microtubules/metabolism,ultrastructure Molecular Weight Ovum/enzymology Sea Urchins
Chemicals
Adenosine Triphosphate Adenosine Triphosphatases Dyneins
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Hollenbeck P J
Suprynowicz F
Cande W Z
References (42)
42 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.99.4.1251
Published
1984-10-00
Pages
1251-8
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2113311
Subset
IM
Grants
NIGMS NIH HHS · GM23238 · United States
External Links
PubMed source DOI: 10.1083/jcb.99.4.1251 PMC Full Text View journal details
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