Experimental Characterization of the Interaction between the N-Terminal SH3 Domain of Crkl and C3G.
Adaptor Proteins, Signal Transducing
/ chemistry
Binding Sites
Cell Adhesion
Cell Differentiation
Cell Proliferation
Guanine Nucleotide-Releasing Factor 2
/ chemistry
Humans
Models, Molecular
Mutagenesis, Site-Directed
/ methods
Protein Binding
Protein Conformation
Protein Domains
Static Electricity
kinetics
site-directed mutagenesis
stopped-flow
Journal
International journal of molecular sciences
ISSN: 1422-0067
Titre abrégé: Int J Mol Sci
Pays: Switzerland
ID NLM: 101092791
Informations de publication
Date de publication:
07 Dec 2021
07 Dec 2021
Historique:
received:
27
10
2021
revised:
25
11
2021
accepted:
03
12
2021
entrez:
24
12
2021
pubmed:
25
12
2021
medline:
3
2
2022
Statut:
epublish
Résumé
Crkl is a protein involved in the onset of several cancer pathologies that exerts its function only through its protein-protein interaction domains, a SH2 domain and two SH3 domains. SH3 domains are small protein interaction modules that mediate the binding and recognition of proline-rich sequences. One of the main physiological interactors of Crkl is C3G (also known as RAPGEF1), an interaction with key implications in regulating cellular growth and differentiation, cell morphogenesis and adhesion processes. Thus, understanding the interaction between Crkl and C3G is fundamental to gaining information about the molecular determinants of the several cancer pathologies in which these proteins are involved. In this paper, through a combination of fast kinetics at different experimental conditions and site-directed mutagenesis, we characterize the binding reaction between the N-SH3 domain of Crkl and a peptide mimicking a specific portion of C3G. Our results show a clear effect of pH on the stability of the complex, due to the protonation of negatively charged residues in the binding pocket of N-SH3. Our results are discussed under the light of previous work on SH3 domains.
Identifiants
pubmed: 34947971
pii: ijms222413174
doi: 10.3390/ijms222413174
pmc: PMC8705818
pii:
doi:
Substances chimiques
Adaptor Proteins, Signal Transducing
0
CRKL protein
0
Guanine Nucleotide-Releasing Factor 2
0
RAPGEF1 protein, human
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : Sapienza University of Rome
ID : RP11715C34AEAC9B
Organisme : Sapienza University of Rome
ID : RM1181641C2C24B9
Organisme : Sapienza University of Rome
ID : RM11916B414C897E
Organisme : Italian Association for Cancer Research
ID : IG 24551
Organisme : Italian Pasteur Institute
ID : Ariaudo Research Project 2018
Organisme : Marie Skłodowska-Curie UBIMOTIF
ID : 860517
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