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dc.contributor.authorLiaunardy-Jopeace, Ardiyantoen
dc.contributor.authorMurton, Ben Len
dc.contributor.authorMahesh, Mohanen
dc.contributor.authorChin, Jasonen
dc.contributor.authorJames, Johnen
dc.date.accessioned2018-05-03T13:06:02Z
dc.date.available2018-05-03T13:06:02Z
dc.date.issued2017-12en
dc.identifier.issn1545-9993
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/275507
dc.description.abstractLCK is a tyrosine kinase that is essential for initiating T-cell antigen receptor (TCR) signaling. A complete understanding of LCK function is constrained by a paucity of methods to quantitatively study its function within live cells. To address this limitation, we generated LCK*, in which a key active-site lysine is replaced by a photocaged equivalent, using genetic code expansion. This strategy enabled fine temporal and spatial control over kinase activity, thus allowing us to quantify phosphorylation kinetics in situ using biochemical and imaging approaches. We find that autophosphorylation of the LCK active-site loop is indispensable for its catalytic activity and that LCK can stimulate its own activation by adopting a more open conformation, which can be modulated by point mutations. We then show that CD4 and CD8, T-cell coreceptors, can enhance LCK activity, thereby helping to explain their effect in physiological TCR signaling. Our approach also provides general insights into SRC-family kinase dynamics.
dc.format.mediumPrint-Electronicen
dc.languageengen
dc.subjectCD4-Positive T-Lymphocytesen
dc.subjectCD8-Positive T-Lymphocytesen
dc.subjectCell Lineen
dc.subjectHumansen
dc.subjectLymphocyte Specific Protein Tyrosine Kinase p56(lck)en
dc.subjectReceptors, Antigen, T-Cellen
dc.subjectSignal Transductionen
dc.subjectEnzyme Activationen
dc.subjectCatalytic Domainen
dc.subjectPhosphorylationen
dc.subjectZAP-70 Protein-Tyrosine Kinaseen
dc.subjectHEK293 Cellsen
dc.titleEncoding optical control in LCK kinase to quantitatively investigate its activity in live cells.en
dc.typeArticle
prism.endingPage1163
prism.issueIdentifier12en
prism.publicationDate2017en
prism.publicationNameNature structural & molecular biologyen
prism.startingPage1155
prism.volume24en
dc.identifier.doi10.17863/CAM.22741
dcterms.dateAccepted2017-09-25en
rioxxterms.versionofrecord10.1038/nsmb.3492en
rioxxterms.versionAM*
rioxxterms.licenseref.urihttp://www.rioxx.net/licenses/all-rights-reserveden
rioxxterms.licenseref.startdate2017-12en
dc.contributor.orcidChin, Jason [0000-0003-1219-4757]
dc.contributor.orcidJames, John [0000-0003-1452-7578]
dc.identifier.eissn1545-9985
rioxxterms.typeJournal Article/Reviewen
pubs.funder-project-idWellcome Trust (099966/Z/12/Z)


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