File Download

There are no files associated with this item.

  Links for fulltext
     (May Require Subscription)
Supplementary

Article: Examining the mechanism of action of a kinesin inhibitor using stable isotope labeled inhibitors for cross-linking (SILIC)

TitleExamining the mechanism of action of a kinesin inhibitor using stable isotope labeled inhibitors for cross-linking (SILIC)
Authors
Issue Date2011
Citation
Journal of the American Chemical Society, 2011, v. 133, p. 12386-12389 How to Cite?
AbstractIt is difficult to determine a chemical inhibitor’s binding site in multiprotein mixtures, particularly when high-resolution structural studies are not straightforward. Building upon previous research involving photo-cross-linking and the use of mixtures of stable isotopes, we report a method, Stable Isotope Labeled Inhibitors for Cross-linking (SILIC), for mapping a small molecule inhibitor’s binding site in its target protein. In SILIC, structure–activity relationship data is used to design inhibitor analogues that incorporate a photo-cross-linking group along with either natural or ‘heavy’ stable isotopes. An equimolar mixture of these inhibitor analogues is cross-linked to the target protein to yield a robust signature for identifying inhibitor-modified peptide fragments in complex mass spectrometry data. As a proof of concept, we applied this approach to an ATP-competitive inhibitor of kinesin-5, a widely conserved motor protein required for cell division and an anticancer drug target. This analysis, along with mutagenesis studies, suggests that the inhibitor binds at an allosteric site in the motor protein.
Persistent Identifierhttp://hdl.handle.net/10722/222468

 

DC FieldValueLanguage
dc.contributor.authorWacker, S-
dc.contributor.authorKashyap, S-
dc.contributor.authorLi, X-
dc.contributor.authorKapoor, T-
dc.date.accessioned2016-01-18T07:40:59Z-
dc.date.available2016-01-18T07:40:59Z-
dc.date.issued2011-
dc.identifier.citationJournal of the American Chemical Society, 2011, v. 133, p. 12386-12389-
dc.identifier.urihttp://hdl.handle.net/10722/222468-
dc.description.abstractIt is difficult to determine a chemical inhibitor’s binding site in multiprotein mixtures, particularly when high-resolution structural studies are not straightforward. Building upon previous research involving photo-cross-linking and the use of mixtures of stable isotopes, we report a method, Stable Isotope Labeled Inhibitors for Cross-linking (SILIC), for mapping a small molecule inhibitor’s binding site in its target protein. In SILIC, structure–activity relationship data is used to design inhibitor analogues that incorporate a photo-cross-linking group along with either natural or ‘heavy’ stable isotopes. An equimolar mixture of these inhibitor analogues is cross-linked to the target protein to yield a robust signature for identifying inhibitor-modified peptide fragments in complex mass spectrometry data. As a proof of concept, we applied this approach to an ATP-competitive inhibitor of kinesin-5, a widely conserved motor protein required for cell division and an anticancer drug target. This analysis, along with mutagenesis studies, suggests that the inhibitor binds at an allosteric site in the motor protein.-
dc.languageeng-
dc.relation.ispartofJournal of the American Chemical Society-
dc.titleExamining the mechanism of action of a kinesin inhibitor using stable isotope labeled inhibitors for cross-linking (SILIC)-
dc.typeArticle-
dc.identifier.emailLi, X: xiangli@hku.hk-
dc.identifier.authorityLi, X=rp01562-
dc.identifier.doi10.1021/ja204561q-
dc.identifier.hkuros256652-
dc.identifier.volume133-
dc.identifier.spage12386-
dc.identifier.epage12389-

Export via OAI-PMH Interface in XML Formats


OR


Export to Other Non-XML Formats