Interdomain interactions in Grb2 revealed by the conformational stability and CD28 binding analysis.
Circular dichroism
Differential scanning calorimetry
Protein-protein interaction
Size-exclusion chromatography
Surface plasmon resonance
Journal
Biophysical chemistry
ISSN: 1873-4200
Titre abrégé: Biophys Chem
Pays: Netherlands
ID NLM: 0403171
Informations de publication
Date de publication:
05 2022
05 2022
Historique:
received:
30
01
2022
revised:
27
02
2022
accepted:
01
03
2022
pubmed:
12
3
2022
medline:
3
5
2022
entrez:
11
3
2022
Statut:
ppublish
Résumé
Growth-factor receptor-bound protein 2 (Grb2) is an adaptor protein involved in signal transduction. The protein contains a central SH2 domain with N-terminal and C-terminal SH3 domains. We analyzed the SH3 involvement in Grb2 binding to the cytoplasmic region of CD28 receptor, using Grb2 and its SH3-deletion mutants, Grb2_nSH3del and Grb2_cSH3del, as the monomer state. The CD28 binding affinity of Grb2_nSH3del determined from surface plasmon resonance experiments was similar to that of Grb2_cSH3del, which was lower than that of Grb2 and higher than that of Grb2 SH2. The findings indicated that one of the SH3 domains is involved in CD28 binding. We also analyzed the thermal stabilities of Grb2 and its SH3-deletion mutants using differential scanning calorimetry, showing that the order of stability was Grb2_nSH3del < Grb2 < Grb2_cSH3del. Folding thermodynamics results indicated that SH3 domains, particularly nSH3, contribute to stabilizing the structure of Grb2, possibly due to the interdomain interaction.
Identifiants
pubmed: 35276477
pii: S0301-4622(22)00034-5
doi: 10.1016/j.bpc.2022.106792
pii:
doi:
Substances chimiques
CD28 Antigens
0
GRB2 Adaptor Protein
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
106792Informations de copyright
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