1aze
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(New page: 200px<br /> <applet load="1aze" size="450" color="white" frame="true" align="right" spinBox="true" caption="1aze" /> '''NMR STRUCTURE OF THE COMPLEX BETWEEN THE C3...)
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Revision as of 13:57, 12 November 2007
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NMR STRUCTURE OF THE COMPLEX BETWEEN THE C32S-Y7V MUTANT OF THE NSH3 DOMAIN OF GRB2 WITH A PEPTIDE FROM SOS, 10 STRUCTURES
Contents |
Overview
Quantitative analysis of Grb2/dynamin interaction through plasmon, resonance analysis (BIAcore) using Grb2 mutants showed that the high, affinity measured between Grb2 and dynamin is essentially mediated by the, N-SH3 domain of Grb2. In order to study the interactions between Grb2 and, either dynamin or Sos in more detail, Grb2 N-SH3 domains containing, different mutations have been analysed. Two mutations were located on the, hydrophobic platform binding proline-rich peptides (Y7V and P49L) and one, (E40T) located in a region that we had previously shown to be essential, for Grb2/dynamin interactions. Through NMR analysis, we have clearly, demonstrated that the structure of the P49L mutant is not folded, while, the other E40T and Y7V mutants adopt folded structures that are quite, similar to that described for the reference domain. Nevertheless, these, point mutations were shown to alter the overall stability of these domains, by inducing an equilibrium between a folded and an unfolded form. The, complex formed between the peptide VPPPVPPRRR, derived from Sos, and the, E40T mutant was shown to have the same 3D structure as that described for, the wild-type SH3 domain. However, the VPPPVPPRRR peptide adopts a, slightly different orientation when it is complexed with the Y7V mutant., Finally, the affinity of the proline-rich peptide GPPPQVPSRPNR, derived, from dynamin, for the Grb2 N-SH3 domain was too low to be analyzed by NMR., Thus, the interaction between either Sos or dynamin and the SH3 mutants, were tested on a cellular homogenate by means of a far-Western blot, analysis. In these conditions, the P49L mutant was shown to be devoid of, affinity for Sos as well as for dynamin. The Y7V SH3 mutant displayed a, decrease of affinity for both Sos and dynamin, while the E40T mutant, exhibited a decrease of affinity only for dynamin. These results support, the existence of two binding sites between dynamin and the Grb2 N-SH3, domain.
Disease
Known diseases associated with this structure: Central hypoventilation syndrome, congenital OMIM:[100790], Haddad syndrome OMIM:[100790]
About this Structure
1AZE is a Protein complex structure of sequences from Drosophila melanogaster and Homo sapiens. Full crystallographic information is available from OCA.
Reference
Molecular and cellular analysis of Grb2 SH3 domain mutants: interaction with Sos and dynamin., Vidal M, Goudreau N, Cornille F, Cussac D, Gincel E, Garbay C, J Mol Biol. 1999 Jul 16;290(3):717-30. PMID:10395825
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