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

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

21 Scopus citations

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 (US)
Pages (from-to)	1257-1262
Number of pages	6
Journal	ACS Synthetic Biology
Volume	6
Issue number	7
DOIs	https://doi.org/10.1021/acssynbio.6b00359
State	Published - Jul 21 2017

All Science Journal Classification (ASJC) codes

Biomedical Engineering
Biochemistry, Genetics and Molecular Biology (miscellaneous)

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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.",

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.}",

note = "Publisher Copyright: {\textcopyright} 2017 American Chemical Society.",

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doi = "10.1021/acssynbio.6b00359",

language = "English (US)",

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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.

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Engineering Pak1 Allosteric Switches

Abstract

All Science Journal Classification (ASJC) codes

UN SDGs

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