Source:http://linkedlifedata.com/resource/pubmed/id/12866878
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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
6
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| pubmed:dateCreated |
2003-7-17
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| pubmed:abstractText |
Analysis of protein complexes using hydrogen exchange (HX) combined with high resolution electrospray mass spectrometry (MS) is demonstrated. HX MS offers the possibility to analyze the strength of binding in protein complexes, to identify regions that undergo binding induced structural changes, and to study the nature (hydrophobic, electrostatic, etc.) of binding between two or more proteins. In the current work, a heteromeric complex containing UBC9 (an E2 conjugating enzyme) and SUMO-1 (a ubiquitin-like modifier) was investigated by incubating the complex in D2O and measuring the amount of deuterium incorporation with MS. SUMO-1 had significant changes in deuterium levels when bound to UBC9. In contract, few or no changes in deuterium levels were detected in UBC9 when part of the complex, even at the binding interface. Titrations were used to estimate the binding constant for the complex. The nature of the interface was probed by creating a site-directed mutant form of UBC9. The mutant form showed no detectable binding to SUMO-1 and thereby suggested that binding between these two proteins is primarily electrostatically driven. This application of HX MS demonstrates its value in the study of protein complexes and protein machinery.
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| pubmed:language |
eng
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| pubmed:journal | |
| pubmed:citationSubset |
IM
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| pubmed:chemical | |
| pubmed:status |
MEDLINE
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| pubmed:month |
Jun
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| pubmed:issn |
0003-2654
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| pubmed:author | |
| pubmed:issnType |
Print
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| pubmed:volume |
128
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| pubmed:owner |
NLM
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| pubmed:authorsComplete |
Y
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| pubmed:pagination |
623-8
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| pubmed:dateRevised |
2004-11-17
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| pubmed:meshHeading |
pubmed-meshheading:12866878-Escherichia coli,
pubmed-meshheading:12866878-Humans,
pubmed-meshheading:12866878-Hydrogen,
pubmed-meshheading:12866878-Ion Exchange,
pubmed-meshheading:12866878-Protein Binding,
pubmed-meshheading:12866878-Proteins,
pubmed-meshheading:12866878-SUMO-1 Protein,
pubmed-meshheading:12866878-Spectrometry, Mass, Electrospray Ionization
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| pubmed:year |
2003
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| pubmed:articleTitle |
Analysis of protein complexes with hydrogen exchange and mass spectrometry.
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| pubmed:affiliation |
Department of Chemistry, Clark Hall 242 MSC03-2060, University of New Mexico, Albuquerque, NM 87131-0001, USA. engen@unm.edu
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| pubmed:publicationType |
Journal Article
|