Source:http://linkedlifedata.com/resource/pubmed/id/11327818
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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
6
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| pubmed:dateCreated |
2001-4-30
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| pubmed:abstractText |
Cytoplasmic dynein is a multisubunit ATPase that transforms chemical energy into motion along microtubules. LC8, a 10 kDa light chain subunit of the dynein complex, is highly conserved with 94% sequence identity between Drosophila and human. The precise function of this protein is unknown, but its ubiquitous expression and conservation suggest a critical role in the function of the dynein motor complex. We have overexpressed LC8 from Drosophila melanogaster and characterized its dimerization and folding using analytical ultracentrifugation, size-exclusion chromatography, circular dichroism, and fluorescence spectroscopy. Sedimentation equilibrium measurements of LC8 at pH 7 reveal a reversible monomer-dimer equilibrium with a dissociation constant of 12 microM at 4 degrees C. At lower pH, LC8 dissociates to a monomer, with a transition midpoint at pH 4.8. Far-UV CD and fluorescence spectra demonstrate that pH-dissociated LC8 retains native secondary and tertiary structures, while the diminished near-UV CD signal shows loss of quaternary structure. The observation that dimeric LC8 dissociates at low pH can be explained by titration of a histidine pair in the dimer interface. Equilibrium denaturation experiments with a protein concentration range spanning almost 2 orders of magnitude indicate that unfolding of LC8 dimer is a two-stage process, in which global unfolding is preceded by dissociation to a folded monomer. The nativelike tertiary structure of the monomer suggests a role for the monomer-dimer equilibrium of LC8 in dynein function.
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| pubmed:grant | |
| pubmed:language |
eng
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| pubmed:journal | |
| pubmed:citationSubset |
IM
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| pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Carrier Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Drosophila Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Dyneins,
http://linkedlifedata.com/resource/pubmed/chemical/Fungal Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Guanidine,
http://linkedlifedata.com/resource/pubmed/chemical/Insect Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/ctp protein, Drosophila
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| pubmed:status |
MEDLINE
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| pubmed:month |
Feb
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| pubmed:issn |
0006-2960
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| pubmed:author | |
| pubmed:issnType |
Print
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| pubmed:day |
13
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| pubmed:volume |
40
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| pubmed:owner |
NLM
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| pubmed:authorsComplete |
Y
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| pubmed:pagination |
1596-605
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| pubmed:dateRevised |
2009-12-11
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| pubmed:meshHeading |
pubmed-meshheading:11327818-Animals,
pubmed-meshheading:11327818-Aspergillus nidulans,
pubmed-meshheading:11327818-Carrier Proteins,
pubmed-meshheading:11327818-Chromatography, Gel,
pubmed-meshheading:11327818-Circular Dichroism,
pubmed-meshheading:11327818-Conserved Sequence,
pubmed-meshheading:11327818-Cytoplasm,
pubmed-meshheading:11327818-Dimerization,
pubmed-meshheading:11327818-Drosophila Proteins,
pubmed-meshheading:11327818-Drosophila melanogaster,
pubmed-meshheading:11327818-Dyneins,
pubmed-meshheading:11327818-Fungal Proteins,
pubmed-meshheading:11327818-Guanidine,
pubmed-meshheading:11327818-Humans,
pubmed-meshheading:11327818-Insect Proteins,
pubmed-meshheading:11327818-Protein Conformation,
pubmed-meshheading:11327818-Protein Denaturation,
pubmed-meshheading:11327818-Protein Folding,
pubmed-meshheading:11327818-Spectrometry, Fluorescence,
pubmed-meshheading:11327818-Thermodynamics,
pubmed-meshheading:11327818-Ultracentrifugation
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| pubmed:year |
2001
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| pubmed:articleTitle |
Dimerization and folding of LC8, a highly conserved light chain of cytoplasmic dynein.
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| pubmed:affiliation |
Department of Chemistry and Biochemistry, Ohio University, Athens, OH 45701, USA. barbar@ohio.edu
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| pubmed:publicationType |
Journal Article,
Comparative Study,
Research Support, U.S. Gov't, P.H.S.,
Research Support, Non-U.S. Gov't
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