Autophosphorylation is a mechanism of inhibition in twitchin kinase
Autophosphorylation is a mechanism of inhibition in twitchin kinase
No Thumbnail Available
Files
There are no files associated with this item.
Date
2018
Authors
Wilkinson, Mark
Rigden, Daniel J.
Benian, Guy M.
Eyers, Patrick A.
Editors
Journal ISSN
Electronic ISSN
ISBN
Bibliographical data
Publisher
Series
DOI (citable link)
International patent number
Link to the license
oops
EU project number
Project
Open Access publication
Collections
Title in another language
Publication type
Journal article
Publication status
Published
Published in
Journal of molecular biology ; 430 (2018), 6. - pp. 793-805. - ISSN 0022-2836. - eISSN 1089-8638
Abstract
Titin-like kinases are muscle-specific kinases that regulate mechanical sensing in the sarcomere. Twitchin kinase (TwcK) is the best-characterized member of this family, both structurally and enzymatically. TwcK activity is auto-inhibited by a dual intrasteric mechanism, in which N- and C-terminal tail extensions wrap around the kinase domain, blocking the hinge region, the ATP binding pocket and the peptide substrate binding groove. Physiologically, kinase activation is thought to occur by a stretch-induced displacement of the inhibitory tails from the kinase domain. Here, we now show that TwcK inhibits its catalysis even in the absence of regulatory tails, by undergoing auto-phosphorylation at mechanistically important elements of the kinase fold. Using mass spectrometry, site-directed mutagenesis and catalytic assays on recombinant samples we identify residues T212, T301, T316 and T401 as primary auto-phosphorylation sites in TwcK in vitro. Taken together, our results suggest that residue T316, located in the peptide substrate binding P+1 loop, is the dominantly regulatory site in TwcK. Based on these findings, we conclude that TwcK is regulated through a triple inhibitory mechanism consisting of phosphorylation as well as intrasteric blockage, which is responsive not only to mechanical cues but also to biochemical modulation. This implies that mechanically stretched conformations of TwcK do not necessarily correspond to catalytically active states, as previously postulated. This further suggests a phosphorylation-dependent desensitization of the TwcK-mediated mechanoresponse of the sarcomere in vivo.
Summary in another language
Subject (DDC)
570 Biosciences, Biology
Keywords
Kinase regulation; mass spectrometry; site-directed mutagenesis; phosphotransfer catalysis
Conference
Review
undefined / . - undefined, undefined. - (undefined; undefined)
Cite This
ISO 690
WILLIAMS, Rhys, Barbara FRANKE, Mark WILKINSON, Jennifer S. FLEMING, Daniel J. RIGDEN, Guy M. BENIAN, Patrick A. EYERS, Olga MAYANS, 2018. Autophosphorylation is a mechanism of inhibition in twitchin kinase. In: Journal of molecular biology. 430(6), pp. 793-805. ISSN 0022-2836. eISSN 1089-8638. Available under: doi: 10.1016/j.jmb.2018.01.020BibTex
@article{Williams2018-03Autop-41302, year={2018}, doi={10.1016/j.jmb.2018.01.020}, title={Autophosphorylation is a mechanism of inhibition in twitchin kinase}, number={6}, volume={430}, issn={0022-2836}, journal={Journal of molecular biology}, pages={793--805}, author={Williams, Rhys and Franke, Barbara and Wilkinson, Mark and Fleming, Jennifer S. and Rigden, Daniel J. and Benian, Guy M. and Eyers, Patrick A. and Mayans, Olga} }
RDF
<rdf:RDF xmlns:dcterms="http://purl.org/dc/terms/" xmlns:dc="http://purl.org/dc/elements/1.1/" xmlns:rdf="http://www.w3.org/1999/02/22-rdf-syntax-ns#" xmlns:bibo="http://purl.org/ontology/bibo/" xmlns:dspace="http://digital-repositories.org/ontologies/dspace/0.1.0#" xmlns:foaf="http://xmlns.com/foaf/0.1/" xmlns:void="http://rdfs.org/ns/void#" xmlns:xsd="http://www.w3.org/2001/XMLSchema#" > <rdf:Description rdf:about="https://kops.uni-konstanz.de/server/rdf/resource/123456789/41302"> <dc:contributor>Eyers, Patrick A.</dc:contributor> <dc:creator>Mayans, Olga</dc:creator> <dc:contributor>Mayans, Olga</dc:contributor> <dc:creator>Benian, Guy M.</dc:creator> <dc:contributor>Rigden, Daniel J.</dc:contributor> <dc:creator>Rigden, Daniel J.</dc:creator> <dc:contributor>Franke, Barbara</dc:contributor> <dc:creator>Eyers, Patrick A.</dc:creator> <dc:creator>Williams, Rhys</dc:creator> <void:sparqlEndpoint rdf:resource="http://localhost/fuseki/dspace/sparql"/> <dcterms:title>Autophosphorylation is a mechanism of inhibition in twitchin kinase</dcterms:title> <dcterms:isPartOf rdf:resource="https://kops.uni-konstanz.de/server/rdf/resource/123456789/28"/> <foaf:homepage rdf:resource="http://localhost:8080/"/> <dc:creator>Wilkinson, Mark</dc:creator> <dc:contributor>Fleming, Jennifer S.</dc:contributor> <dc:contributor>Benian, Guy M.</dc:contributor> <dc:creator>Franke, Barbara</dc:creator> <dc:contributor>Wilkinson, Mark</dc:contributor> <dcterms:issued>2018-03</dcterms:issued> <dcterms:abstract xml:lang="eng">Titin-like kinases are muscle-specific kinases that regulate mechanical sensing in the sarcomere. Twitchin kinase (TwcK) is the best-characterized member of this family, both structurally and enzymatically. TwcK activity is auto-inhibited by a dual intrasteric mechanism, in which N- and C-terminal tail extensions wrap around the kinase domain, blocking the hinge region, the ATP binding pocket and the peptide substrate binding groove. Physiologically, kinase activation is thought to occur by a stretch-induced displacement of the inhibitory tails from the kinase domain. Here, we now show that TwcK inhibits its catalysis even in the absence of regulatory tails, by undergoing auto-phosphorylation at mechanistically important elements of the kinase fold. Using mass spectrometry, site-directed mutagenesis and catalytic assays on recombinant samples we identify residues T212, T301, T316 and T401 as primary auto-phosphorylation sites in TwcK in vitro. Taken together, our results suggest that residue T316, located in the peptide substrate binding P+1 loop, is the dominantly regulatory site in TwcK. Based on these findings, we conclude that TwcK is regulated through a triple inhibitory mechanism consisting of phosphorylation as well as intrasteric blockage, which is responsive not only to mechanical cues but also to biochemical modulation. This implies that mechanically stretched conformations of TwcK do not necessarily correspond to catalytically active states, as previously postulated. This further suggests a phosphorylation-dependent desensitization of the TwcK-mediated mechanoresponse of the sarcomere in vivo.</dcterms:abstract> <dcterms:available rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2018-02-13T12:35:20Z</dcterms:available> <dc:contributor>Williams, Rhys</dc:contributor> <dc:creator>Fleming, Jennifer S.</dc:creator> <dc:language>eng</dc:language> <bibo:uri rdf:resource="https://kops.uni-konstanz.de/handle/123456789/41302"/> <dc:date rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2018-02-13T12:35:20Z</dc:date> <dspace:isPartOfCollection rdf:resource="https://kops.uni-konstanz.de/server/rdf/resource/123456789/28"/> </rdf:Description> </rdf:RDF>
Internal note
xmlui.Submission.submit.DescribeStep.inputForms.label.kops_note_fromSubmitter
Examination date of dissertation
Method of financing
Comment on publication
Alliance license
Corresponding Authors der Uni Konstanz vorhanden
International Co-Authors
Bibliography of Konstanz
Yes
Refereed
Yes