Document Detail

Engineering a lever into the kinesin neck.
MedLine Citation:
PMID:  9792635     Owner:  NLM     Status:  MEDLINE    
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.
M Mazumdar; R A Cross
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  273     ISSN:  0021-9258     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  1998 Nov 
Date Detail:
Created Date:  1998-12-10     Completed Date:  1998-12-10     Revised Date:  2003-11-14    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  29352-9     Citation Subset:  IM    
Molecular Motors Group, Marie Curie Research Institute, The Chart, Oxted, Surrey RH8 0TL, United Kingdom.
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MeSH Terms
Actinin / genetics
Amino Acid Sequence
Base Sequence
DNA Primers
Kinesin / chemistry,  genetics*
Molecular Sequence Data
Mutagenesis, Insertional
Protein Engineering
Reg. No./Substance:
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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