Maydan, M;
McDonald, PC;
Sanghera, J;
Yan, J;
Rallis, C;
Pinchin, S;
Hannigan, GE;
... Dedhar, S; + view all
(2010)
Integrin-Linked Kinase Is a Functional Mn2+-Dependent Protein Kinase that Regulates Glycogen Synthase Kinase-3 beta (GSK-3 beta) Phosphorylation.
PLOS ONE
, 5
(8)
, Article e12356. 10.1371/journal.pone.0012356.
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Abstract
Background: Integrin-linked kinase (ILK) is a highly evolutionarily conserved, multi-domain signaling protein that localizes to focal adhesions, myofilaments and centrosomes where it forms distinct multi-protein complexes to regulate cell adhesion, cell contraction, actin cytoskeletal organization and mitotic spindle assembly. Numerous studies have demonstrated that ILK can regulate the phosphorylation of various protein and peptide substrates in vitro, as well as the phosphorylation of potential substrates and various signaling pathways in cultured cell systems. Nevertheless, the ability of ILK to function as a protein kinase has been questioned because of its atypical kinase domain.Methodology/Principal Findings: Here, we have expressed full-length recombinant ILK, purified it to >94% homogeneity, and characterized its kinase activity. Recombinant ILK readily phosphorylates glycogen synthase kinase-3 (GSK-3) peptide and the 20-kDa regulatory light chains of myosin (LC20). Phosphorylation kinetics are similar to those of other active kinases, and mutation of the ATP-binding lysine (K220 within subdomain 2) causes marked reduction in enzymatic activity. We show that ILK is a Mn-dependent kinase (the K-m for MnATP is similar to 150-fold less than that for MgATP).Conclusions/Significance: Taken together, our data demonstrate that ILK is a bona fide protein kinase with enzyme kinetic properties similar to other active protein kinases.
| Type: | Article |
|---|---|
| Title: | Integrin-Linked Kinase Is a Functional Mn2+-Dependent Protein Kinase that Regulates Glycogen Synthase Kinase-3 beta (GSK-3 beta) Phosphorylation |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.1371/journal.pone.0012356 |
| Publisher version: | http://dx.doi.org/10.1371/journal.pone.0012356 |
| Language: | English |
| Additional information: | © 2010 Maydan et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. This work was supported by grants from the Canadian Cancer Society Research Institute (SD), Cancer Research UK (DIH) and the Canadian Institutes of Health Research (MPW). SignalChem Inc (JS and JY) produced and provided the recombinant proteins used in the study. As such, they were involved in data collection and preparation of the manuscript as it relates to the production of these recombinant proteins. The remaining funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. |
| Keywords: | PHOSPHATASE TARGET SUBUNIT, TUMOR-SUPPRESSOR PTEN, BINDING-PROTEIN, BETA-CATENIN, DEPENDENT PATHWAYS, ENDOTHELIAL-CELLS, B/AKT ACTIVATION, FOCAL ADHESIONS, ILK, SURVIVAL |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences > Cell and Developmental Biology |
| URI: | https://discovery.ucl.ac.uk/id/eprint/142439 |
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