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PDBsum entry 2btt
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Contractile protein
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PDB id
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2btt
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References listed in PDB file
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Key reference
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Title
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New approaches to high-Throughput structure characterization of sh3 complexes: the example of myosin-3 and myosin-5 sh3 domains from s. Cerevisiae.
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Authors
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V.Musi,
B.Birdsall,
G.Fernandez-Ballester,
R.Guerrini,
S.Salvatori,
L.Serrano,
A.Pastore.
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Ref.
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Protein Sci, 2006,
15,
795-807.
[DOI no: ]
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PubMed id
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Abstract
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SH3 domains are small protein modules that are involved in protein-protein
interactions in several essential metabolic pathways. The availability of the
complete genome and the limited number of clearly identifiable SH3 domains make
the yeast Saccharomyces cerevisae an ideal proteomic-based model system to
investigate the structural rules dictating the SH3-mediated protein interactions
and to develop new tools to assist these studies. In the present work, we have
determined the solution structure of the SH3 domain from Myo3 and modeled by
homology that of the highly homologous Myo5, two myosins implicated in actin
polymerization. We have then implemented an integrated approach that makes use
of experimental and computational methods to characterize their binding
properties. While accommodating their targets in the classical groove, the two
domains have selectivity in both orientation and sequence specificity of the
target peptides. From our study, we propose a consensus sequence that may
provide a useful guideline to identify new natural partners and suggest a
strategy of more general applicability that may be of use in other structural
proteomic studies.
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Figure 3.
Figure 3 Relaxation parameters for Myo3-SH3 (left panel)
and Myo5-SH3 (right panel) recorded at 600 MHz and 20°C. R1
and R2 and ^15N-NOE are plotted as a function of residue number.
The values for the amide resonances of residues 19, 56, 57, and
64–66 for Myo3-SH3 and 2, 3, 18, 19, 57, 59, and 61 for
Myo5-SH3 are missing because they were not assigned, whereas the
other missing values correspond to residues that were excluded
either because of overlap or because they were too weak to be
properly appreciated.
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Figure 5.
Figure 5 Effects of the peptide titrations on the Myo-SH3
NMR spectra. A representative example of Myo3-SH3 titrated with
3BP-1 is shown. (A) Superposition of the HSQC spectra recorded
at 25°C and 500 MHz corresponding to 0, 0.33, 1, and 3 molar
Myo3-SH3:peptide molar ratios colored in black, red, green, and
blue, respectively. (B) Histogram of the weighted chemical
shifted variation as observed for a 1:3 molar ratio of
Myo3-SH3:3BP-1 complex. (C) Mapping of the interaction surface
of 3BP-1 onto the structure of Myo3-SH3. The secondary structure
elements are indicated with red and yellow (the  -helix) and
cyan (the  -sheet) solid
oval ribbons. The side chains of residues with amide resonances
most dramatically shifted upon binding of the peptide are
explicitly shown.
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The above figures are
reprinted
by permission from the Protein Society:
Protein Sci
(2006,
15,
795-807)
copyright 2006.
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