Structural and biophysical properties of farnesylated KRas interacting with the chaperone SmgGDS-558.
Journal
Biophysical journal
ISSN: 1542-0086
Titre abrégé: Biophys J
Pays: United States
ID NLM: 0370626
Informations de publication
Date de publication:
04 10 2022
04 10 2022
Historique:
received:
07
02
2022
revised:
02
05
2022
accepted:
19
05
2022
pubmed:
27
5
2022
medline:
12
10
2022
entrez:
26
5
2022
Statut:
ppublish
Résumé
KRas is a small GTPase and membrane-bound signaling protein. Newly synthesized KRas is post-translationally modified with a membrane-anchoring prenyl group. KRas chaperones are therapeutic targets in cancer due to their participation in trafficking oncogenic KRas to membranes. SmgGDS splice variants are chaperones for small GTPases with basic residues in their hypervariable domain (HVR), including KRas. SmgGDS-607 escorts pre-prenylated small GTPases, while SmgGDS-558 escorts prenylated small GTPases. We provide a structural description of farnesylated and fully processed KRas (KRas-FMe) in complex with SmgGDS-558 and define biophysical properties of this interaction. Surface plasmon resonance measurements on biomimetic model membranes quantified the thermodynamics of the interaction of SmgGDS with KRas, and small-angle x-ray scattering was used to characterize complexes of SmgGDS-558 and KRas-FMe structurally. Structural models were refined using Monte Carlo and molecular dynamics simulations. Our results indicate that SmgGDS-558 interacts with the HVR and the farnesylated C-terminus of KRas-FMe, but not its G-domain. Therefore, SmgGDS-558 interacts differently with prenylated KRas than prenylated RhoA, whose G-domain was found in close contact with SmgGDS-558 in a recent crystal structure. Using immunoprecipitation assays, we show that SmgGDS-558 binds the GTP-bound, GDP-bound, and nucleotide-free forms of farnesylated and fully processed KRas in cells, consistent with SmgGDS-558 not engaging the G-domain of KRas. We found that the dissociation constant, K
Identifiants
pubmed: 35614853
pii: S0006-3495(22)00421-0
doi: 10.1016/j.bpj.2022.05.028
pmc: PMC9617131
pii:
doi:
Substances chimiques
Guanine Nucleotide Exchange Factors
0
Molecular Chaperones
0
Guanosine Triphosphate
86-01-1
Monomeric GTP-Binding Proteins
EC 3.6.5.2
Types de publication
Journal Article
Research Support, U.S. Gov't, Non-P.H.S.
Research Support, N.I.H., Extramural
Langues
eng
Sous-ensembles de citation
IM
Pagination
3684-3697Subventions
Organisme : NCI NIH HHS
ID : R01 CA188871
Pays : United States
Informations de copyright
Copyright © 2022 Biophysical Society. All rights reserved.
Déclaration de conflit d'intérêts
Declaration of interests The authors declare no competing interests.
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