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PDBsum entry 2w10
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References listed in PDB file
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Key reference
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Title
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Distinct binding modes of two epitopes in gab2 that interact with the sh3c domain of grb2.
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Authors
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M.Harkiolaki,
T.Tsirka,
M.Lewitzky,
P.C.Simister,
D.Joshi,
L.E.Bird,
E.Y.Jones,
N.O'Reilly,
S.M.Feller.
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Ref.
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Structure, 2009,
17,
809-822.
[DOI no: ]
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PubMed id
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Abstract
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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.
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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.
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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.
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The above figures are
reprinted
by permission from Cell Press:
Structure
(2009,
17,
809-822)
copyright 2009.
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