| Engineering a lever into the kinesin neck. | |
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MedLine Citation:
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PMID: 9792635 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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To probe for a lever arm action in the kinesin stepping mechanism, we engineered a rodlike extension piece into the tail of rat kinesin at various points close to the head-tail junction and measured its effects on the temperature dependence of velocity in microtubule gliding assays. The insert comprised two contiguous alpha-actinin triple-coil repeats and was predicted to fold into a stiff rodlike module about 11 nm long. The effects of this module were greater the closer it was placed to the head-tail junction. When inserted distal to the head-tail junction, at Asn401 in the dimeric K partial differential401GST, the insert had no effect. When inserted closer to the heads at Val376 into K partial differential376GST, the insert slowed progress below 22 degreesC but accelerated progress to approximately 125% of wild type above 22 degreesC. The most dramatic effect of the synthetic lever occurred when it was inserted very close to the head-neck junction, at Glu340 into the single-headed construct K partial differential340GST. This construct was immotile without the insert, but motile with it, at about 30% of the velocity of the dimeric control. The alpha-actinin module thus confers some gain-of-function when inserted close to the head-neck junction but not when placed distal to it. The data exclude the presence of a lever arm C-terminal to Val376 in the kinesin tail but suggest that a short-throw lever arm may be present, N-terminal to Val376 and contiguous with the head-neck junction at Ala339. |
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Authors:
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M Mazumdar; R A Cross |
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Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: The Journal of biological chemistry Volume: 273 ISSN: 0021-9258 ISO Abbreviation: J. Biol. Chem. Publication Date: 1998 Nov |
Date Detail:
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Created Date: 1998-12-10 Completed Date: 1998-12-10 Revised Date: 2003-11-14 |
Medline Journal Info:
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Nlm Unique ID: 2985121R Medline TA: J Biol Chem Country: UNITED STATES |
Other Details:
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Languages: eng Pagination: 29352-9 Citation Subset: IM |
Affiliation:
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Molecular Motors Group, Marie Curie Research Institute, The Chart, Oxted, Surrey RH8 0TL, United Kingdom. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Actinin
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genetics Amino Acid Sequence Animals Base Sequence DNA Primers Kinesin / chemistry, genetics* Molecular Sequence Data Mutagenesis, Insertional Protein Engineering Rats |
| Chemical | |
Reg. No./Substance:
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0/DNA Primers; 11003-00-2/Actinin; EC 3.6.1.-/Kinesin |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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