Show simple item record

dc.contributor.authorDodson, CAen_US
dc.contributor.authorBayliss, Ren_US
dc.date.accessioned2018-08-13T10:37:53Z
dc.date.issued2012-01-06en_US
dc.identifier2en_US
dc.identifier.citationJOURNAL OF BIOLOGICAL CHEMISTRY, 2012, 287 pp. 1150 - 1157en_US
dc.identifier.issn0021-9258en_US
dc.identifier.urihttps://repository.icr.ac.uk/handle/internal/2316
dc.identifier.doi10.1074/jbc.M111.312090en_US
dc.description.abstractProtein kinases are activated by phosphorylation and by the binding of activator proteins. The interplay of these two factors is incompletely understood. We applied energetic analysis to this question and characterized the activation process of the serine/threonine kinase Aurora-A by phosphorylation and by its protein partner, targeting protein for Xenopus kinesin-like protein 2 (TPX2). We discovered that these two activators act synergistically and without a predefined order: each can individually increase the activity of Aurora-A, and the effect of both bound together is the exact sum of their individual contributions to catalysis. Unexpectedly, the unphosphorylated enzyme has catalytic activity that is increased 15-fold by the binding of TPX2 alone. The energetic contribution of phosphorylation to catalysis is 2-fold greater than that of TPX2 binding, which is independent of the phosphorylation state of the enzyme. Based on this analysis, we propose a revised, fluid model of Aurora-A activation in which the first step is a reduction in the mobility of the activation loop by either TPX2 binding or phosphorylation. Furthermore, our results suggest that unphosphorylated Aurora-A bound to the mitotic spindle by TPX2 is catalytically active and that the phosphorylation state of Aurora-A is an inaccurate surrogate for its activity. Extending this form of analysis will allow us to compare quantitatively the effects of the whole network of kinase-activating partners. Comparison with other kinases showed that kinetic characterization detects those kinases whose activation loops undergo a rearrangement upon phosphorylation and thus whose unphosphorylated state offers a distinct target for the development of Type II inhibitors.en_US
dc.format.extent1150 - 1157en_US
dc.languageEnglishen_US
dc.language.isoEnglishen_US
dc.publisherAMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INCen_US
dc.rights.urihttp://www.rioxx.net/licenses/all-rights-reserveden_US
dc.titleActivation of Aurora-A Kinase by Protein Partner Binding and Phosphorylation Are Independent and Synergisticen_US
dc.typeJournal Article
rioxxterms.versionofrecord10.1074/jbc.M111.312090en_US
rioxxterms.licenseref.startdate2012-01-06en_US
rioxxterms.typeJournal Article/Reviewen_US
dc.relation.isPartOfJOURNAL OF BIOLOGICAL CHEMISTRYen_US
pubs.notesaffiliation: Bayliss, R (Reprint Author), Univ Leicester, Dept Biochem, Lancaster Rd, Leicester LE1 9HN, Leics, England. Dodson, Charlotte A.; Bayliss, Richard, Inst Canc Res, Div Struct Biol, London SW3 6JB, England. keywords-plus: MITOTIC SPINDLE; RESISTANT AURORA; STRUCTURAL BASIS; KINETIC BASIS; INHIBITORS; TPX2; MECHANISM; DISCOVERY; SUBSTRATE; MUTANTS research-areas: Biochemistry & Molecular Biology web-of-science-categories: Biochemistry & Molecular Biology author-email: [email protected] orcid-numbers: Bayliss, Richard/0000-0003-0604-2773 Dodson, Charlotte/0000-0002-8403-726X funding-acknowledgement: Royal Society; Cancer Research UK [C24461/A8032]; Institute of Cancer Research; National Health Service funding-text: This work was supported by a Royal Society University Research Fellowship (to R. B.) and Cancer Research UK Project Grant C24461/A8032 (to R. B.).; Mass spectrometry data acquisition and analysis were carried out by Angela Paul and Andrew Thompson at The Institute of Cancer Research Proteomics Core Facility. We thank Jessica Schmitt, Kathy Boxall, Wynne Aherne, Rosemary Burke, and Rob van Montfort in the Division of Cancer Therapeutics for practical advice and use of the Caliper EZ Reader system; Tim Sharpe (Department of Biochemistry, University of Cambridge) for helpful discussions; Mark Richards (The Institute of Cancer Research) for mutagenesis; and Clare Stace (The Institute of Cancer Research) for protein production. We acknowledge the Career Development Faculty Programme of The Institute of Cancer Research and National Health Service funding to the National Institute for Health Research Biomedical Research Centre. number-of-cited-references: 26 times-cited: 46 usage-count-last-180-days: 0 usage-count-since-2013: 9 journal-iso: J. Biol. Chem. doc-delivery-number: 877IE unique-id: ISI:000299170300030 oa: gold_or_bronze da: 2018-08-13en_US
pubs.notesNot knownen_US
pubs.organisational-group/ICR
pubs.volume287en_US
pubs.embargo.termsNot knownen_US
dc.contributor.icrauthorDodson, Charlotteen_US
dc.contributor.icrauthorBayliss, Richarden_US


Files in this item

Thumbnail

This item appears in the following Collection(s)

Show simple item record