Reduced dynamic complexity allows structure elucidation of an excited state of KRASG13D

Chao, Fa-An; Chan, Albert H.; Dharmaiah, Srisathiyanarayanan; Schwieters, Charles D.; Tran, Timothy H.; Taylor, Troy; Ramakrishnan, Nitya; Esposito, Dominic; Nissley, Dwight V.; McCormick, Frank; Simanshu, Dhirendra K.; Cornilescu, Gabriel

doi:10.1038/s42003-023-04960-6

Reduced dynamic complexity allows structure elucidation of an excited state of KRAS^G13D

Journal Article · Fri Jun 02 00:00:00 EDT 2023 · Communications Biology

DOI:https://doi.org/10.1038/s42003-023-04960-6· OSTI ID:2423582

Chao, Fa-An ^[1]; Chan, Albert H. ^[1]; ^[1]; ^[2]; Tran, Timothy H. ^[1]; Taylor, Troy ^[1]; Ramakrishnan, Nitya ^[1]; ^[1]; ^[1]; ^[3]; ^[1]; ^[1]

Frederick National Laboratory for Cancer Research, Frederick, MD (United States)
National Institutes of Health (NIH), Bethesda, MD (United States)
Frederick National Laboratory for Cancer Research, Frederick, MD (United States); Univ. of California, San Francisco, CA (United States)

Localized dynamics of RAS, including regions distal to the nucleotide-binding site, is of high interest for elucidating the mechanisms by which RAS proteins interact with effectors and regulators and for designing inhibitors. Among several oncogenic mutants, methyl relaxation dispersion experiments reveal highly synchronized conformational dynamics in the active (GMPPNP-bound) KRAS^G13D, which suggests an exchange between two conformational states in solution. Methyl and ³¹P NMR spectra of active KRAS^G13D in solution confirm a two-state ensemble interconverting on the millisecond timescale, with a major Pγ atom peak corresponding to the dominant State 1 conformation and a secondary peak indicating an intermediate state different from the known State 2 conformation recognized by RAS effectors. High-resolution crystal structures of active KRAS^G13D and KRAS^G13D-RAF1 RBD complex provide snapshots of the State 1 and 2 conformations, respectively. We use residual dipolar couplings to solve and cross-validate the structure of the intermediate state of active KRAS^G13D, showing a conformation distinct from those of States 1 and 2 outside the known flexible switch regions. The dynamic coupling between the conformational exchange in the effector lobe and the breathing motion in the allosteric lobe is further validated by a secondary mutation in the allosteric lobe, which affects the conformational population equilibrium.

View Accepted Manuscript (DOE)

Research Organization:: Argonne National Laboratory (ANL), Argonne, IL (United States). Advanced Photon Source (APS)

Sponsoring Organization:: National Institutes of Health (NIH); USDOE Office of Science (SC), Basic Energy Sciences (BES). Scientific User Facilities (SUF)

Grant/Contract Number:: AC02-06CH11357

OSTI ID:: 2423582

Journal Information:: Communications Biology, Journal Name: Communications Biology Journal Issue: 1 Vol. 6; ISSN 2399-3642

Publisher:: Springer NatureCopyright Statement

Country of Publication:: United States

Language:: English

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