Journal article
A robust methodology to subclassify pseudokinases based on their nucleotide-binding properties
JM Murphy, Q Zhang, SN Young, ML Reese, FP Bailey, PA Eyers, D Ungureanu, H Hammaren, O Silvennoinen, LN Varghese, K Chen, A Tripaydonis, N Jura, K Fukuda, J Qin, Z Nimchuk, MB Mudgett, S Elowe, CL Gee, L Liu Show all
Biochemical Journal | Published : 2014
DOI: 10.1042/BJ20131174
Abstract
Protein kinase-like domains that lack conserved residues known to catalyse phosphoryl transfer, termed pseudokinases, have emerged as important signalling domains across all kingdoms of life. Although predicted to function principally as catalysisindependent protein-interaction modules, several pseudokinase domains have been attributed unexpected catalytic functions, often amid controversy. We established a thermal-shift assay as a benchmark technique to define the nucleotide-binding properties of kinase-like domains. Unlike in vitro kinase assays, this assay is insensitive to the presence of minor quantities of contaminating kinases thatmay otherwise lead to incorrect attribution of catalyt..
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Awarded by Medical Research Council
Funding Acknowledgements
This work was supported by the National Health and Medical Research Council (NHMRC) [grant numbers 637342 and 10118041; the Australian Research Council (ARC) [fellowships FT100100100 and FT110100169 (to J.M.M. and J.J.B.)]; the Leukaemia Foundation and the Australian Stem Cell Centre via scholarships to L.N.V.; the National Institutes of Health (NIH) [grant numbers R01 HL58758 (to J.Q.) and AI73756 (to M.L.R.)]; the Medical Research Council of Academy of Finland, the Sigrid Juselius Foundation, the Medical Research Fund of Tampere University Hospital, the Finnish Cancer Foundation and the Tampere Tuberculosis Foundation (to O.S.); the American Heart Association [grant number 11BGIA7440051 (to N.J.)]; and the National Science Foundation [grant number IOS-0821801 (to M.B.M.)]; with additional support from the Victorian State Government Operational Infrastructure Support and the NHMRC Independent Research Institutes Infrastructure Support Scheme (IRIISS) [grant number 361646].