title: Sequence determinants of a specific inactive protein kinase conformation.
abstract: Only a small percentage of protein kinases have been shown to adopt a distinct inactive ATP-binding site conformation, called the Asp-Phe-Gly-out (DFG-out) conformation. Given the high degree of homology within this enzyme family, we sought to understand the basis of this disparity on a sequence level. We identified two residue positions that sensitize mitogen-activated protein kinases (MAPKs) to inhibitors that stabilize the DFG-out inactive conformation. After characterizing the structure and dynamics of an inhibitor-sensitive MAPK mutant, we demonstrated the generality of this strategy by sensitizing a kinase (apoptosis signal-regulating kinase 1) not in the MAPK family to several DFG-out stabilizing ligands, using the same residue positions. The use of specific inactive conformations may aid the study of noncatalytic roles of protein kinases, such as binding partner interactions and scaffolding effects.
authors.affiliation: Department of Chemistry, University of Washington, Seattle, WA 98195, USA.
authors.forename: Sanjay B
authors.initials: SB
authors.lastname: Hari
authors.forename: Ethan A
authors.initials: EA
authors.lastname: Merritt
authors.forename: Dustin J
authors.initials: DJ
authors.lastname: Maly
mesh_terms: D020816:Amino Acid Motifs; D020134:Catalytic Domain; D007700:Kinetics; D008024:Ligands; D048848:MAP Kinase Kinase Kinase 5 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D019950:Mitogen-Activated Protein Kinase 1 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D020928:Mitogen-Activated Protein Kinases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015335:Molecular Probes / Q000737:chemistry / Q000378:metabolism; D009154:Mutation; D011485:Protein Binding; D047428:Protein Kinase Inhibitors / Q000737:chemistry / Q000378:metabolism; D017434:Protein Structure, Tertiary
chemical_list: D008024:Ligands; D015335:Molecular Probes; D047428:Protein Kinase Inhibitors; D019950:Mitogen-Activated Protein Kinase 1; D020928:Mitogen-Activated Protein Kinases; D048848:MAP Kinase Kinase Kinase 5