title: Structural investigations of a GYF domain covalently linked to a proline-rich peptide.
abstract: Protein structure determination of low affinity complexes of interacting macromolecules is often hampered by a lack of observable NOEs between the binding partners. Covalent linkage offers a way to shift the equilibrium of the interaction partners to the bound state. Here we show that a single-chain protein containing the GYF domain of CD2BP2 and the target peptide SHRPPPPGHRV from CD2 allows for the intramolecular association of the binding partners. We obtained NOEs between the GYF domain and the peptide that could define the principal orientation of the peptide in the complex. In conjunction with general recognition rules for proline-rich sequence recognition these NOEs allowed the accurate modeling of the protein-peptide complex.
authors.affiliation: Protein Engineering Group, Forschungsinstitut für Molekulare Pharmakologie and Freie Universität Berlin, Robert-Rössle-Strasse 10, 13125 Berlin, Germany. freund@fmp-berlin.de
authors.forename: Christian
authors.initials: C
authors.lastname: Freund
authors.forename: Ronald
authors.initials: R
authors.lastname: Kühne
authors.forename: Sunghyouk
authors.initials: S
authors.lastname: Park
authors.forename: Katharina
authors.initials: K
authors.lastname: Thiemke
authors.forename: Ellis L
authors.initials: EL
authors.lastname: Reinherz
authors.forename: Gerhard
authors.initials: G
authors.lastname: Wagner
mesh_terms: D009682:Magnetic Resonance Spectroscopy; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism; D011392:Proline / Q000378:metabolism; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism
chemical_list: D010455:Peptides; D011506:Proteins; D011392:Proline