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PMID: 6147357 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Taxol-induced bundling of brain-derived microtubules.

The Journal of cell biology ·Vol. 99 ·No. 3 ·1984-09-00 ·Pages 940-6

Turner PF, Margolis RL

Abstract

Taxol has two obvious effects in cells. It stabilizes microtubules and it induces microtubule bundling. We have duplicated the microtubule-bundling effect of taxol in vitro and report preliminary characterization of this bundling using electron microscopy, sedimentation, and electrophoretic analyses. Taxol-bundled microtubules from rat brain crude extracts were seen as massive bundles by electron microscopy. Bundled microtubules sedimented through sucrose five times faster than control microtubules. Electrophoretic analysis of control and taxol-bundled microtubules pelleted through sucrose revealed no striking differences between the two samples except for a protein doublet of approximately 100,000 daltons. Taxol-induced microtubule bundling was not produced by using pure tubulin or recycled microtubule protein; this suggested that taxol-induced microtubule bundling was mediated by a factor present in rat brain crude extracts. Taxol cross-linked rat brain crude extract microtubules were entirely labile to ATP in the millimolar range. This ATP-dependent relaxation was also demonstrated in a more purified system, using taxol-bundled microtubules pelleted through sucrose and gently resuspended. Although the bundling factor did not recycle with microtubule protein, it was apparently retained on isolated taxol-stabilized microtubules. The bundling factor was salt extracted from taxol-stabilized microtubules and its retained activity was demonstrated in an add-back experiment with assembled phosphocellulose-purified tubulin.

MeSH Terms
Adenosine Triphosphate/pharmacology Alkaloids/pharmacology Animals Brain/ultrastructure Cell Fractionation Centrifugation, Density Gradient Kinetics Microtubule-Associated Proteins Microtubules/drug effects,ultrastructure Nerve Tissue Proteins/metabolism Paclitaxel Proteins/metabolism Rats Rats, Inbred Strains
Chemicals
Alkaloids Microtubule-Associated Proteins Nerve Tissue Proteins Proteins Adenosine Triphosphate Paclitaxel
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Turner P F
Margolis R L
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41 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.99.3.940
Published
1984-09-00
Pages
940-6
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2113414
Subset
IM
Grants
NIEHS NIH HHS · 1 T32 ES07032-05 · United States
NIGMS NIH HHS · GM28189 · United States
NIGMS NIH HHS · GM32022 · United States
External Links
PubMed source DOI: 10.1083/jcb.99.3.940 PMC Full Text View journal details
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