title: Engineering Pak1 Allosteric Switches.
abstract: P21-activated kinases (PAKs) are important regulators of cell motility and morphology. It has been challenging to interrogate their functions because cells adapt to genetic manipulation of PAK, and because inhibitors act on multiple PAK isoforms. Here we describe genetically encoded PAK1 analogues that can be selectively activated by the membrane-permeable small molecule rapamycin. An engineered domain inserted away from the active site responds to rapamycin to allosterically control activity of the PAK1 isoform. To examine the mechanism of rapamycin-induced PAK1 activation, we used molecular dynamics with graph theory to predict amino acids involved in allosteric communication with the active site. This analysis revealed allosteric pathways that were exploited to generate kinase switches. Activation of PAK1 resulted in transient cell spreading in metastatic breast cancer cells, and long-term dendritic spine enlargement in mouse hippocampal CA1 neurons.
authors.forename: Onur
authors.identifier: 0000-0003-1825-1011
authors.initials: O
authors.lastname: Dagliyan
authors.affiliation: Department of Pharmacology, University of Illinois at Chicago , Chicago Illinois 60612, United States.
authors.forename: Andrei V
authors.initials: AV
authors.lastname: Karginov
authors.affiliation: Center for Disease Biology and Integrative Medicine, The University of Tokyo , Bunko-ku, Tokyo 113-0033, Japan.
authors.forename: Sho
authors.initials: S
authors.lastname: Yagishita
authors.affiliation: Division of Cancer Studies, King's College London , London SE1 1UL, England, U.K.
authors.forename: Madeline E
authors.initials: ME
authors.lastname: Gale
authors.forename: Hui
authors.initials: H
authors.lastname: Wang
authors.affiliation: Department of Cell and Molecular Biology, Scripps Research Institute , La Jolla, California 92037, United States.
authors.forename: Celine
authors.initials: C
authors.lastname: DerMardirossian
authors.affiliation: Division of Cancer Studies, King's College London , London SE1 1UL, England, U.K.
authors.forename: Claire M
authors.initials: CM
authors.lastname: Wells
authors.forename: Nikolay V
authors.initials: NV
authors.lastname: Dokholyan
authors.affiliation: Center for Disease Biology and Integrative Medicine, The University of Tokyo , Bunko-ku, Tokyo 113-0033, Japan.
authors.forename: Haruo
authors.initials: H
authors.lastname: Kasai
authors.forename: Klaus M
authors.initials: KM
authors.lastname: Hahn
mesh_terms: D000494:Allosteric Regulation / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D000818:Animals; D056547:CA1 Region, Hippocampal / Q000378:metabolism; D020134:Catalytic Domain / Q000187:drug effects / Q000235:genetics; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects / Q000235:genetics / Q000502:physiology; D006624:Hippocampus / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D051379:Mice; D051381:Rats; D017207:Rats, Sprague-Dawley; D015398:Signal Transduction / Q000187:drug effects / Q000235:genetics / Q000502:physiology; D020123:Sirolimus / Q000494:pharmacology; D054462:p21-Activated Kinases / Q000235:genetics / Q000378:metabolism*
chemical_list: D054462:p21-Activated Kinases; D020123:Sirolimus
keywords: PAK; allosteric switch; cell motility; dendritic spines; protein dynamics