Expression, refolding and crystallizations of the Grb2-like (GADS) C-terminal SH3 domain complexed with a SLP-76 motif peptide.
The Grb2-like adaptor protein GADS is composed of an N-terminal SH3 domain, an SH2 domain, a proline-rich region and a C-terminal SH3 domain. GADS interacts through its C-terminal SH3 domain with the adaptor protein SLP-76, thus recruiting this protein and other associated molecules to the linker for activation of T-cell (LAT) protein. The DNA encoding the C-terminal SH3 domain of GADS (GADS-cSH3) was assembled synthetically using a recursive PCR technique and the protein was overexpressed in Escherichia coli, refolded and purified. Several crystals of this domain in complex with the SLP-76 peptide were obtained and characterized.
Faravelli
Alessandro
A
Istituto Giannina Gaslini, Molecular Medicine, Largo Gerolamo Gaslini 5, 16147 Genova, Italy.
Dimasi
Nazzareno
N
D048868:Adaptor Proteins, Signal Transducing / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D020816:Amino Acid Motifs; D000818:Animals; D003460:Crystallization; D018360:Crystallography, X-Ray; D051379:Mice; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry* / Q000235:genetics; D010750:Phosphoproteins / Q000737:chemistry*; D017510:Protein Folding; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D018909:src Homology Domains / Q000235:genetics
D048868:Adaptor Proteins, Signal Transducing; C498726:Mona protein, mouse; D010446:Peptide Fragments; D010750:Phosphoproteins; D011994:Recombinant Proteins; C092947:SLP-76 signal Transducing adaptor proteins