Engineering Pak1 Allosteric Switches

Onur Dagliyan; Andrei V. Karginov; Sho Yagishita; Madeline E. Gale; Hui Wang; Celine Dermardirossian; Claire M. Wells; Nikolay V. Dokholyan; Haruo Kasai; Klaus M. Hahn

doi:10.1021/acssynbio.6b00359

Engineering Pak1 Allosteric Switches

Onur Dagliyan, Andrei V. Karginov, Sho Yagishita, Madeline E. Gale, Hui Wang, Celine Dermardirossian, Claire M. Wells, Nikolay V. Dokholyan, Haruo Kasai, Klaus M. Hahn^*

^*Corresponding author for this work

Research output: Contribution to journal › Letter › peer-review

26 Citations (Scopus)

217 Downloads (Pure)

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.

Original language	English
Pages (from-to)	1257-1262
Number of pages	6
Journal	ACS synthetic biology
Volume	6
Issue number	7
Early online date	3 Apr 2017
DOIs	https://doi.org/10.1021/acssynbio.6b00359
Publication status	Published - 21 Jul 2017

Keywords

allosteric switch
cell motility
dendritic spines
PAK
protein dynamics

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1021/acssynbio.6b00359

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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.",

keywords = "allosteric switch, cell motility, dendritic spines, PAK, protein dynamics",

author = "Onur Dagliyan and Karginov, {Andrei V.} and Sho Yagishita and Gale, {Madeline E.} and Hui Wang and Celine Dermardirossian and Wells, {Claire M.} and Dokholyan, {Nikolay V.} and Haruo Kasai and Hahn, {Klaus M.}",

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doi = "10.1021/acssynbio.6b00359",

language = "English",

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TY - JOUR

T1 - Engineering Pak1 Allosteric Switches

AU - Dagliyan, Onur

AU - Karginov, Andrei V.

AU - Yagishita, Sho

AU - Gale, Madeline E.

AU - Wang, Hui

AU - Dermardirossian, Celine

AU - Wells, Claire M.

AU - Dokholyan, Nikolay V.

AU - Kasai, Haruo

AU - Hahn, Klaus M.

PY - 2017/7/21

Y1 - 2017/7/21

N2 - 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.

AB - 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.

KW - allosteric switch

KW - cell motility

KW - dendritic spines

KW - PAK

KW - protein dynamics

UR - http://www.scopus.com/inward/record.url?scp=85025121891&partnerID=8YFLogxK

U2 - 10.1021/acssynbio.6b00359

DO - 10.1021/acssynbio.6b00359

M3 - Letter

AN - SCOPUS:85025121891

SN - 2161-5063

VL - 6

SP - 1257

EP - 1262

JO - ACS synthetic biology

JF - ACS synthetic biology

IS - 7

ER -

Engineering Pak1 Allosteric Switches

Abstract

Keywords

UN SDGs

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