title: Sequential assignment of proline-rich regions in proteins: application to modular binding domain complexes.
abstract: Many protein-protein interactions involve amino acid sequences containing proline-rich motifs and even polyproline stretches. The lack of amide protons in such regions complicates assignment, since 1HN-based triple-resonance assignment strategies cannot be employed. Two such systems that we are currently studying include an SH2 domain from the protein Crk with a region containing 9 prolines in a 14 amino acid sequence, as well as a WW domain that interacts with a proline-rich target. A modified version of the HACAN pulse scheme, originally described by Bax and co-workers [Wang et al. (1995) J. Biomol. NMR, 5, 376-382], and an experiment which correlates the intra-residue 1Halpha, 13Calpha/13Cbeta chemical shifts with the 15N shift of the subsequent residue are presented and applied to the two systems listed above, allowing sequential assignment of the molecules.
authors.affiliation: Department of Biochemistry, The University of Toronto, ON, Canada.
authors.forename: V
authors.initials: V
authors.lastname: Kanelis
authors.forename: L
authors.initials: L
authors.lastname: Donaldson
authors.forename: D R
authors.initials: DR
authors.lastname: Muhandiram
authors.forename: D
authors.initials: D
authors.lastname: Rotin
authors.forename: J D
authors.initials: JD
authors.lastname: Forman-Kay
authors.forename: L E
authors.initials: LE
authors.lastname: Kay
mesh_terms: D000595:Amino Acid Sequence; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D011392:Proline / Q000737:chemistry*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000737:chemistry*
chemical_list: D011506:Proteins; D011392:Proline