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A sequential mechanism for clathrin cage disassembly by 70-kDa heat-shock cognate protein (Hsc70) and auxilin
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Rothnie, Alice, Clarke, Anthony R., Kuzmic, Petr, Cameron, Angus and Smith, Corinne J. (2011) A sequential mechanism for clathrin cage disassembly by 70-kDa heat-shock cognate protein (Hsc70) and auxilin. National Academy of Sciences. Proceedings, Vol.108 (No.17). pp. 6927-6932. doi:10.1073/pnas.1018845108 ISSN 0027-8424.
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Official URL: http://dx.doi.org/10.1073/pnas.1018845108
Abstract
An essential stage in endocytic coated vesicle recycling is the dissociation of clathrin from
the vesicle coat by the molecular chaperone, Hsc70 and the J-domain-containing protein,
auxilin, in an ATP-dependent process. We present the first detailed mechanistic analysis
of clathrin disassembly catalysed by Hsc70 and auxilin, using loss of perpendicular light
scattering to monitor the process. We report that a single auxilin per clathrin triskelion is
required for maximal rate of disassembly, that ATP is hydrolysed at the same rate that
disassembly occurs and that three ATP molecules are hydrolysed per clathrin triskelion
released. Stopped-flow measurements revealed a lag phase in which the scattering
intensity increased owing to association of Hsc70 with clathrin cages followed by serial
rounds of ATP hydrolysis prior to triskelion removal. Global fit of stopped flow data to
several physically plausible mechanisms showed the best fit to a model in which
sequential hydrolysis of three separate ATP molecules is required for the eventual
release of a triskelion from the clathrin-auxilin cage.
Item Type: | Journal Article | ||||
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Subjects: | Q Science > QH Natural history | ||||
Divisions: | Faculty of Science, Engineering and Medicine > Science > Life Sciences (2010- ) > Biological Sciences ( -2010) Faculty of Science, Engineering and Medicine > Science > Life Sciences (2010- ) |
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Library of Congress Subject Headings (LCSH): | Coated vesicles, Endocytosis, Adenosine triphosphate | ||||
Journal or Publication Title: | National Academy of Sciences. Proceedings | ||||
Publisher: | National Academy of Sciences | ||||
ISSN: | 0027-8424 | ||||
Official Date: | 26 April 2011 | ||||
Dates: |
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Volume: | Vol.108 | ||||
Number: | No.17 | ||||
Page Range: | pp. 6927-6932 | ||||
DOI: | 10.1073/pnas.1018845108 | ||||
Status: | Peer Reviewed | ||||
Publication Status: | Published | ||||
Access rights to Published version: | Restricted or Subscription Access | ||||
Funder: | Medical Research Council (Great Britain) (MRC), Wellcome Trust (London, England) | ||||
Grant number: | G0601125 (MRC), 055663/Z/98/Z (WT) |
Data sourced from Thomson Reuters' Web of Knowledge
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