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Cited by 95Microtubule and mitosis dynamicsGenetic and Kidney Cyst DiseasesProtein Structure and Dynamics

Isolation of a 45-kDa fragment from the kinesin heavy chain with enhanced ATPase and microtubule-binding activities

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Kuznetsov, S A, Vaisberg, Y A, Rothwell, S W, Murphy, D B, Gelfand, V I·1989·Journal of Biological Chemistry
DOI 10.1016/s0021-9258(17)31301-7
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Abstract

Kinesin is a microtubule-activated, mechanochemical ATPase capable of moving particles along microtubules and making microtubules glide along a solid substrate. In this study we used limited proteolysis to study the structure of bovine brain kinesin, a heterotetramer composed of two heavy (120-kDa) and two light (62-kDa) chains. alpha-chymotrypsin, trypsin, and subtilisin all produced a protease-resistant 45-kDa fragment from the kinesin heavy chain. As isolated by gel-filtration chromatography, this fragment contains both the microtubule-binding site and the ATP catalytic site of the molecule. Proteolytic cleavage stimulated microtubule-dependent Mg2+-ATPase activity 4- to 5-fold up to 75-120 mumol ATP/min/mg. Cleavage also increased the affinity of the fragment for microtubules at least 10-fold. Since the purified fragment does not support the gliding of flagellar axonemes, we propose that cleavage of the heavy chain uncouples ATPase activity from its translocator activity, which may require other parts of the molecule.

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  • Soluble kinesin purified from bovine brain has only moderate ATPase activity upon interaction with MTs, but the ATPase activity is much higher after proteolytic removal of the COOH-terminal half of the molecule.

    “Soluble kinesin purified from bovine brain has only a moderate ATPase activity upon interaction with MTs, but the ATPase activity is much higher after proteolytic removal of the COOH-terminal half of the molecule (Kuznetsov et al., 1989).”

    From Light Chain– dependent Regulation of Kinesin's Interaction with Microtubules (1998) · cites this paper as Kuznetsov et al., 1989

    The Abstract reports that proteolytic cleavage of bovine brain kinesin increased microtubule-dependent ATPase activity, and the Discussion describes progressive activation as a 45-kDa heavy-chain fragment appears. The passages identify that fragment as part of the heavy chain but do not establish that the removed portion was specifically its COOH-terminal half. Page numbers were not supplied in our copy. (LLM, conf=0.86)

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  • Inhibition of motor activity by the HC tail is in agreement with previous reports that the motor domain alone has higher MT-stimulated ATPase activity than intact kinesin.

    “Inhibition of motor activity by the HC tail is in agreement with previous reports that the motor domain alone has higher MT-stimulated ATPase activity than intact kinesin (Kuznetsov et al., 1989; Gilbert and Johnson, 1993; Stewart et al., 1993; Huang and Hackney, 1994; discussed in Hackney, 1996).”

    From Light Chain– dependent Regulation of Kinesin's Interaction with Microtubules (1998) · cites this paper as Kuznetsov et al., 1989

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Cites

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Research works printed but not linked: 3

  1. Kuznetsov, S. A., and Gelfand, V. I. (1986) Proc. Natl. Acad. Sci.
  2. Kuznetsov, S. A., Vaisberg, E. A., Shanina, N. A., Magretova, N. N., Chernyak, V. Y., and Gelfand, V. I. (1988) EMBO J. 7 , Shelanski, M. L., Gaskin, F., and Cantor, C. R. (1973) Proc. Natl. Acad. Sci. U. S. A. 70,765-768
  3. Weingarten, M. D., Lockwood, A. H., Hwo, S. Y., and Kirschner, M. W. (1975) Proc. Natl. Acad. Sci. U. S. A . 7 5 , 1858-1862

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Authors
Kuznetsov, S. A., Vaisberg, Y. A., Rothwell, S. W., Murphy, D. B., & Gelfand, V. I.
Affiliations
Johns Hopkins University (US)
ORCID
Venue
Journal of Biological Chemistry
Publisher
Elsevier BV
Published
1989-01
Volume
264
Issue
1
Pages
589-595
ISSN
0021-9258 (ISSN-L)
DOI
10.1016/s0021-9258(17)31301-7
OpenAlex
W1487975069
Cited by
95
References
38
FWCI
5.20
Type
journal article
Language
en
Open access
Licence
Locations
  • Journal of Biological Chemistry (publishedVersion, cc-by)
  • PubMed (publishedVersion)
Funders
  • National Institutes of Health
  • National Institute of General Medical Sciences (GM33171)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Microtubule and mitosis dynamics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Genetic and Kidney Cyst Diseases
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
Keywords
kinesin, kinesin heavy chain, ATPase activity, microtubule-binding site, proteolytic cleavage, limited proteolysis, microtubule gliding, flagellar axoneme, translocation, bovine brain
MeSH terms
Adenosine Triphosphatases, Animals, Brain, Cattle, Chymotrypsin, Kinetics, Microtubule Proteins, Microtubules, Molecular Weight, Nerve Tissue Proteins, Peptide Fragments, Protein Binding, Tubulin, Kinesins

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