Controlling the Activity of the Tec Kinase Itk by Mutation of the Phenylalanine Gatekeeper Residue

dc.contributor.author Joseph, Raji
dc.contributor.author Andreotti, Amy
dc.contributor.author Andreotti, Amy
dc.contributor.department Biochemistry, Biophysics and Molecular Biology
dc.date 2018-02-17T03:47:08.000
dc.date.accessioned 2020-06-29T23:46:24Z
dc.date.available 2020-06-29T23:46:24Z
dc.date.copyright Sat Jan 01 00:00:00 UTC 2011
dc.date.issued 2010-12-01
dc.description.abstract <p>The regulatory spine is a set of conserved residues that are assembled and disassembled upon activation and inactivation of kinases. We recently identified the regulatory spine within the immunologically important Tec family kinases and have shown that in addition to the core spine residues within the kinase domain itself, contributions from the SH2−kinase linker region result in an extended spine structure for this kinase family. Disruption of the regulatory spine, either by mutation or by removal of the amino-terminal SH2−kinase linker region or by mutation of core spine residues, leads to inactivation of the Tec kinases. With a focus on the Tec family members, Itk and Btk, we now show that the gatekeeper residue is also critical for the assembly of the regulatory spine. Mutation of the bulky Itk F434 gatekeeper residue to alanine or glycine inactivates Itk. The activity of the Itk F434A mutant can be recovered by a secondary site mutation within the N-terminal lobe, specifically L432I. The Itk L432I mutation likely rescues the activity of the gatekeeper F434A mutation by promoting the assembly of the regulatory spine. We also show that mutation of the Itk and Btk gatekeeper residues to methionine is sufficient to activate the isolated kinase domains of Tec kinases in the absence of the amino-terminal SH2−kinase linker. Thus, shifting the conformational equilibrium between the assembled and disassembled states of the regulatory spine by changing the nature of the gatekeeper residue is key to regulating the activity of Tec kinases.</p>
dc.description.comments <p>Reprinted (adapted) with permission from <em>Biochemistry </em>50 (2011): 221, doi:<a href="http://dx.doi.org/10.1021/bi101379m" target="_blank">10.1021/bi101379m</a>. Copyright 2011 American Chemical Society.</p>
dc.format.mimetype application/pdf
dc.identifier archive/lib.dr.iastate.edu/bbmb_ag_pubs/20/
dc.identifier.articleid 1025
dc.identifier.contextkey 7747276
dc.identifier.s3bucket isulib-bepress-aws-west
dc.identifier.submissionpath bbmb_ag_pubs/20
dc.identifier.uri https://dr.lib.iastate.edu/handle/20.500.12876/10666
dc.language.iso en
dc.source.bitstream archive/lib.dr.iastate.edu/bbmb_ag_pubs/20/0-AndreottiLicense_ControllingActivity.pdf|||Fri Jan 14 22:17:34 UTC 2022
dc.source.bitstream archive/lib.dr.iastate.edu/bbmb_ag_pubs/20/2011_Andreotti_ControllingActivity.pdf|||Fri Jan 14 22:17:35 UTC 2022
dc.source.uri 10.1021/bi101379m
dc.subject.disciplines Biochemistry, Biophysics, and Structural Biology
dc.subject.disciplines Molecular Biology
dc.title Controlling the Activity of the Tec Kinase Itk by Mutation of the Phenylalanine Gatekeeper Residue
dc.type article
dc.type.genre article
dspace.entity.type Publication
relation.isAuthorOfPublication 97143b55-8496-4365-8699-e254a8ac0903
relation.isOrgUnitOfPublication c70f85ae-e0cd-4dce-96b5-4388aac08b3f
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