SH3 domains of Grb2 adaptor bind to PXψPXR motifs within the Sos1 nucleotide exchange factor in a discriminate manner

Caleb B. McDonald, Kenneth L. Seldeen, Brian J. Deegan, Amjad Farooq

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

Ubiquitously encountered in a wide variety of cellular processes, the Grb2-Sos1 interaction is mediated through the combinatorial binding of nSH3 and cSH3 domains of Grb2 to various sites containing PXψPXR motifs within Sos1. Here, using isothermal titration calorimetry, we demonstrate that while the nSH3 domain binds with affinities in the physiological range to all four sites containing PXψPXR motifs, designated S1, S2, S3, and S4, the cSH3 domain can only do so at the S1 site. Further scrutiny of these sites yields rationale for the recognition of various PXψPXR motifs by the SH3 domains in a discriminate manner. Unlike the PXψPXR motifs at S2, S3, and S4 sites, the PXψPXR motif at the S1 site is flanked at its C-terminus with two additional arginine residues that are absolutely required for high-affinity binding of the cSH3 domain. In striking contrast, these two additional arginine residues augment the binding of the nSH3 domain to the S1 site, but their role is not critical for the recognition of S2, S3, and S4 sites. Site-directed mutagenesis suggests that the two additional arginine residues flanking the PXψPXR motif at the S1 site contribute to free energy of binding via the formation of salt bridges with specific acidic residues in SH3 domains. Molecular modeling is employed to project these novel findings into the 3D structures of SH3 domains in complex with a peptide containing the PXψPXR motif and flanking arginine residues at the S1 site. Taken together, this study furthers our understanding of the assembly of a key signaling complex central to cellular machinery.

Original languageEnglish
Pages (from-to)4074-4085
Number of pages12
JournalBiochemistry
Volume48
Issue number19
DOIs
StatePublished - May 19 2009

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src Homology Domains
Arginine
Nucleotides
Mutagenesis
Calorimetry
Molecular modeling
Site-Directed Mutagenesis
Titration
Free energy
Machinery
Salts
Peptides

ASJC Scopus subject areas

  • Biochemistry

Cite this

SH3 domains of Grb2 adaptor bind to PXψPXR motifs within the Sos1 nucleotide exchange factor in a discriminate manner. / McDonald, Caleb B.; Seldeen, Kenneth L.; Deegan, Brian J.; Farooq, Amjad.

In: Biochemistry, Vol. 48, No. 19, 19.05.2009, p. 4074-4085.

Research output: Contribution to journalArticle

McDonald, Caleb B. ; Seldeen, Kenneth L. ; Deegan, Brian J. ; Farooq, Amjad. / SH3 domains of Grb2 adaptor bind to PXψPXR motifs within the Sos1 nucleotide exchange factor in a discriminate manner. In: Biochemistry. 2009 ; Vol. 48, No. 19. pp. 4074-4085.
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