1fi6
From Proteopedia
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SOLUTION STRUCTURE OF THE REPS1 EH DOMAIN
Overview
The recently described EH domain recognizes proteins containing, Asn-Pro-Phe (NPF) sequences. Using nuclear magnetic resonance (NMR) data, we determined the solution structure of the EH domain from the Reps1, protein and characterized its binding to linear and cyclic peptides, derived from a novel targeting protein. The structure calculation included, 1143 distance restraints and 122 angle restraints and resulted in, structures with a root-mean-square deviation of 0.40 +/- 0.05 A for, backbone atoms of superimposed secondary structural elements. The, structure comprises two helix-loop-helix motifs characteristic of EF-hand, domains. Titration data with NPF-containing peptides showed evidence of, intermediate exchange on the NMR chemical shift time scale, which required, an analysis that includes curve fitting to obtain accurate equilibrium, constants and dissociation rate constants. The cyclic and linear peptides, bound with similar affinities (Kd = 65 +/- 17 and 46 +/- 14 microM, respectively) and to the same hydrophobic pocket formed between helices B, and C. The cyclic peptide formed a complex that dissociated more slowly, (k(off) = 440 +/- 110 s(-1)) than the linear peptide (k(off) = 1800 +/-, 250 s(-1)), but had little change in affinity because of the slower rate, of association of the cyclic peptide. In addition, we characterized, binding to a peptide containing a DPF sequence (Kd = 0.5 +/- 0.2 mM). The, characterization of binding between the Reps1 EH domain and its target, proteins provides information about their role in endocytosis.
About this Structure
1FI6 is a Single protein structure of sequence from Mus musculus with CA as ligand. Full crystallographic information is available from OCA.
Reference
Solution structure of the Reps1 EH domain and characterization of its binding to NPF target sequences., Kim S, Cullis DN, Feig LA, Baleja JD, Biochemistry. 2001 Jun 12;40(23):6776-85. PMID:11389591
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