Engineering Pak1 Allosteric Switches.
Onur DagliyanAndrei V KarginovSho YagishitaMadeline E GaleHui WangCeline DerMardirossianClaire M WellsNikolay V DokholyanHaruo KasaiKlaus M HahnPublished in: ACS synthetic biology (2017)
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.
Keyphrases
- small molecule
- molecular dynamics
- single cell
- breast cancer cells
- squamous cell carcinoma
- small cell lung cancer
- spinal cord
- protein protein
- induced apoptosis
- density functional theory
- spinal cord injury
- escherichia coli
- transcription factor
- gene expression
- stem cells
- cell proliferation
- molecular docking
- pseudomonas aeruginosa
- bone marrow
- dna methylation
- cystic fibrosis
- cell cycle arrest
- oxidative stress
- subarachnoid hemorrhage
- high glucose
- stress induced
- mesenchymal stem cells
- tyrosine kinase
- molecular dynamics simulations
- biofilm formation
- convolutional neural network