title: Label transfer reagents to probe p38 MAPK binding partners.
abstract: Protein kinases are essential enzymes for cellular signaling, and are often regulated by participation in protein complexes. The mitogen-activated protein kinase (MAPK) p38 is involved in multiple pathways, and its regulation depends on its interactions with other signaling proteins. However, the identification of p38-interacting proteins is challenging. For this reason, we have developed label transfer reagents (LTRs) that allow labeling of p38 signaling complexes. These LTRs leverage the potency and selectivity of known p38 inhibitors to place a photo-crosslinker and tag in the vicinity of p38 and its binding partners. Upon UV irradiation, proteins that are in close proximity to p38 are covalently crosslinked, and labeled proteins are detected and/or purified with an orthogonal chemical handle. Here we demonstrate that p38-selective LTRs selectively label a diversity of p38 binding partners, including substrates, activators, and inactivators. Furthermore, these LTRs can be used in immunoprecipitations to provide low-resolution structural information on p38-containing complexes.
authors: Department of Chemistry, University of Washington, Box 351700, Seattle, WA 98195-1700, USA.
authors: Simeon S
authors: SS
authors: Andrews
authors: Zachary B
authors: ZB
authors: Hill
authors: B Gayani K
authors: BG
authors: Perera
authors: Dustin J
authors: DJ
authors: Maly
mesh_terms: D000818:Animals; D002460:Cell Line; D006801:Humans; D007202:Indicators and Reagents; D008958:Models, Molecular; D025941:Protein Interaction Mapping / Q000379:methods*; D015398:Signal Transduction; D013194:Staining and Labeling; D014466:Ultraviolet Rays; D048051:p38 Mitogen-Activated Protein Kinases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D007202:Indicators and Reagents; D048051:p38 Mitogen-Activated Protein Kinases