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“DFG-Flip” in the Insulin Receptor Kinase Is Facilitated by a Helical Intermediate State of the Activation Loop
Journal article   Open access   Peer reviewed

“DFG-Flip” in the Insulin Receptor Kinase Is Facilitated by a Helical Intermediate State of the Activation Loop

Harish Vashisth, Luca Maragliano and Cameron F Abrams
Biophysical journal, v 102(8), pp 1979-1987
18 Apr 2012
PMID: 22768955
url
https://doi.org/10.1016/j.bpj.2012.03.031View
Published, Version of Record (VoR) Open

Abstract

Protein
We have characterized a large-scale inactive-to-active conformational change in the activation-loop of the insulin receptor kinase domain at the atomistic level via untargeted temperature-accelerated molecular dynamics (TAMD) and free-energy calculations using the string method. TAMD simulations consistently show folding of the A-loop into a helical conformation followed by unfolding to an active conformation, causing the highly conserved DFG-motif (Asp 1150 , Phe 1151 , and Gly 1152 ) to switch from the inactive “D-out/F-in” to the nucleotide-binding-competent “D-in/F-out” conformation. The minimum free-energy path computed from the string method preserves these helical intermediates along the inactive-to-active path, and the thermodynamic free-energy differences are consistent with previous work on various other kinases. The mechanisms revealed by TAMD also suggest that the regulatory spine can be dynamically assembled/disassembled either by DFG-flip or by movement of the α C-helix. Together, these findings both broaden our understanding of kinase activation and point to intermediates as specific therapeutic targets.

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Web of Science research areas
Biophysics
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