PDBsum entry 2vvk

Protein binding

PDB id: 2vvk

Contents

Protein chain

56 a.a. *

Ligands

GOL

Waters ×77

* Residue conservation analysis

PDB id:

2vvk

Name:

Protein binding

Title:

Grb2 sh3c (1)

Structure:

Growth factor receptor-bound protein 2. Chain: a. Fragment: sh3 domain, residues 161-214. Synonym: adapter protein grb2, sh2/sh3 adapter grb2, protein ash, grb2 sh3c. Engineered: yes. Other_details: n terminal gs overhand due to thrombin cleavage

Source:

Homo sapiens. Human. Organism_taxid: 9606. Expressed in: escherichia coli. Expression_system_taxid: 469008.

Resolution:

1.60Å R-factor: 0.160 R-free: 0.178

Authors:

M.Harkiolaki,T.Tsirka,S.M.Feller

Key ref:

M.Harkiolaki et al. (2009). Distinct binding modes of two epitopes in Gab2 that interact with the SH3C domain of Grb2. Structure, 17, 809-822. PubMed id: 19523899 DOI: 10.1016/j.str.2009.03.017

Date:

09-Jun-08 Release date: 19-May-09

Protein chain

P62993  (GRB2_HUMAN) -  Growth factor receptor-bound protein 2 from Homo sapiens

Seq: 217 a.a.

Struc: 56 a.a.*

* PDB and UniProt seqs differ at 2 residue positions (black crosses)

Enzyme reactions

• Enzyme class: E.C.?

DOI no: 10.1016/j.str.2009.03.017

Structure 17:809-822 (2009)

PubMed id: 19523899

Distinct binding modes of two epitopes in Gab2 that interact with the SH3C domain of Grb2.

M.Harkiolaki, T.Tsirka, M.Lewitzky, P.C.Simister, D.Joshi, L.E.Bird, E.Y.Jones, N.O'Reilly, S.M.Feller.

ABSTRACT

Grb2 and Gab2 form a complex implicated in normal cell signaling and cancer development. Binding of the Grb2SH3C domain to Gab2 is essential for the interaction, but molecular details remained undefined. Using peptide arrays and isothermal titration calorimetry, two Grb2SH3C binding sites in Gab2 (Gab2a and Gab2b) were confirmed and characterized. Gab2a bears similarity to a p27Kip1 epitope that also binds Grb2SH3C. Crystal structures of both Gab2 epitopes complexed with Grb2SH3C reveal that Gab2b contains a 3(10) helix that positions the arginine and lysine of the core-binding motif RxxK in parallel orientation. In contrast, the Gab2a RxxK motif is embedded in a PPII helix with Arg and Lys in staggered orientation. A similar interaction mode is also present in a new complex of Mona/GadsSH3C with an RxxxxK epitope from the putative phosphatase HD-PTP. In summary, our study reveals interaction types of SH3 domains, highlighting their great versatility.

Selected figure(s)

Figure 1.
Figure 1. Grb2SH3C Domain Overlay Blot of Gab2 Synthesized as Peptide Array Detects Two SH3C-Binding Regions
A schematic drawing of human Gab2 with its N-terminal pleckstrin homology domain (PH) and the five R/KxxK motifs indicated is shown above the peptide array panel. Gab2 was spot-synthesized as overlapping 30 aa peptides, sliding 3 aa with each step. Membrane was initially probed with 2.7 μg/ml GST to detect nonspecific binding, then washed and incubated with anti-GST IgG (rabbit), followed by HRP-coupled anti-rabbit IgG and ECL detection. After visualization of nonspecific signals from the detection system (indicated by dashed boxes), the filter was reblocked and probed with 3.3 μg/ml GST-Grb2SH3C.

Figure 4.
Figure 4. Similarity of the Grb2SH3C-Gab2b Complex with Mona/GadsSH3C in Complex with SLP-76 and HPK1
(A) Alignment of canonical RxxK motifs and structural superposition of the three peptides from Gab2b, SLP-76, and HPK1 at the bound conformations (based on SH3 domain superposition). Electrostatic potential surface representation of (B) Grb2SH3C with Gab2b and Mona/GadsSH3C with (C) SLP-76 and (D) HPK1 peptide bound. Boxed regions in (B), (C), and (D) define the docking position of the RxxK motif on the SH3 domain surface and close-up views of these in atomic detail are provided in panels (E), (F), and (G), respectively.

The above figures are reprinted by permission from Cell Press: Structure (2009, 17, 809-822) copyright 2009.

Figures were selected by an automated process.