Chemical approaches towards unravelling kinase-mediated signalling pathways

Hodgson, David R. W.; Schröder, Martin

doi:10.1039/c0cs00020e

Cited by 18 publications

(18 citation statements)

References 67 publications

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“…Kinase substrates can now be isolated by biochemical or chemical genetic procedures, but both methods require a careful optimization of reaction conditions (Hodgson and Schroder, 2011). For instance, in a chemical genetic approach the typically large hydrophobic or polar residue in the ATP‐binding pocket of the kinase of interest is mutated to a smaller amino acid, such as alanine (A) or glycine (G) (Elphick et al.…”

Section: Discussionmentioning

confidence: 99%

Identification of kinase substrates by bimolecular complementation assays

2012

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SUMMARYAs a consequence of the transient nature of kinase-substrate interactions, the detection of kinase targets, although central for understanding many biological processes, has remained challenging. Here we present a straightforward procedure that relies on the comparison of wild type with activation-loop mutants in the kinase of interest by bimolecular complementation assays. As a proof of functionality, we present the identification and in vivo confirmation of substrates of the major cell-cycle kinase in Arabidopsis, revealing a direct link between cell proliferation and the control of the redox state.

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Section: Discussionmentioning

confidence: 99%

Identification of kinase substrates by bimolecular complementation assays

2012

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“…In the past 10 years, 15 small molecule kinase inhibitors and five anti‐kinase antibodies have been approved by the U.S. Food and Drug Administration (Table 1). These successes (and the projects that failed along the way) have yielded a wealth of reference compounds in the public domain that are useful for investigating the role of specific protein kinases in cellular processes (for reviews, see Shokat and Velleca, 2002; Carlson and White, 2011; Hodgson and Schröder, 2011).…”

Section: The Importance Of Selective Kinase Tool Compoundsmentioning

confidence: 99%

A guide to picking the most selective kinase inhibitor tool compounds for pharmacological validation of drug targets

Uitdehaag

Verkaar

Alwan

et al. 2012

British J Pharmacology

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To establish the druggability of a target, genetic validation needs to be supplemented with pharmacological validation. Pharmacological studies, especially in the kinase field, are hampered by the fact that many reference inhibitors are not fully selective for one target. Fortunately, the initial trickle of selective inhibitors released in the public domain has steadily swelled into a stream. However, rationally picking the most selective tool compound out of the increasing amounts of available inhibitors has become progressively difficult due to the lack of accurate quantitative descriptors of drug selectivity. A recently published approach, termed ‘selectivity entropy’, is an improved way of expressing selectivity as a single‐value parameter and enables rank ordering of inhibitors. We provide a guide to select the best tool compounds for pharmacological validation experiments of candidate drug targets using selectivity entropy. In addition, we recommend which inhibitors to use for studying the biology of the 20 most investigated kinases that are clinically relevant: Abl (ABL1), AKT1, ALK, Aurora A/B, CDKs, MET, CSF1R (FMS), EGFR, FLT3, ERBB2 (HER2), IKBKB (IKK2), JAK2/3, JNK1/2/3 (MAPK8/9/10), MEK1/2, PLK1, PI3Ks, p38α (MAPK14), BRAF, SRC and VEGFR2 (KDR).

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“…We also cover the MS-based conformational analysis approaches for exploring the molecular mechanisms regarding phosphorylation-modulated kinase activation processes, which can also be useful in mapping kinaseinhibitor and kinase-drug interactions for drug discovery efforts. Encouragingly, progresses have been steadily made in the development of substrate database and phosphorylation site-or/and phosphorylation state-specific antibodies as well as in the identification of phosphorylation-dependent protein-protein interactions [108][109][110]. With more advances in phosphoproteomics, bioinformatics, chemical genetics, and chemical biology strategies for functional analysis of protein phosphorylation, we believe that effective integration of these multidisciplinary approaches has made decisive contribution to, and will continue to promote and revive the functional studies on targeted site-specific phosphorylation and their biological roles in cellular signaling processes.…”

Section: Concluding Remarks and Future Perspectivesmentioning

confidence: 99%

Integration of phosphoproteomic, chemical, and biological strategies for the functional analysis of targeted protein phosphorylation

Chen

Huang

2013

Proteomics

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Reversible phosphorylation, tightly controlled by protein kinases and phosphatases, plays a central role in mediating biological processes, such as protein-protein interactions, subcellular translocation, and activation of cellular enzymes. MS-based phosphoproteomics has now allowed the detection and quantification of tens of thousands of phosphorylation sites from a typical biological sample in a single experiment, which has posed new challenges in functional analysis of each and every phosphorylation site on specific signaling phosphoproteins of interest. In this article, we review recent advances in the functional analysis of targeted phosphorylation carried out by various chemical and biological approaches in combination with the MS-based phosphoproteomics. This review focuses on three types of strategies, including forward functional analysis, defined for the result-driven phosphoproteomics efforts in determining the substrates of a specific protein kinase; reverse functional analysis, defined for tracking the kinase(s) for specific phosphosite(s) derived from the discovery-driven phosphoproteomics efforts; and MS-based analysis on the structure-function relationship of phosphoproteins. It is expected that this review will provide a state-of-the-art overview of functional analysis of site-specific phosphorylation and explore new perspectives and outline future challenges.

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Chemical approaches towards unravelling kinase-mediated signalling pathways

Cited by 18 publications

References 67 publications

Identification of kinase substrates by bimolecular complementation assays

Identification of kinase substrates by bimolecular complementation assays

A guide to picking the most selective kinase inhibitor tool compounds for pharmacological validation of drug targets

Integration of phosphoproteomic, chemical, and biological strategies for the functional analysis of targeted protein phosphorylation

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