title: Probing structural and motional features of the C-terminal part of the Human Centrin 2/P17-XPC microcrystalline complex by solid-state NMR spectroscopy.
abstract: Insight into structural and motional features of the C-terminal part of the Human Centrin 2 in complex with the peptide P17-XPC was obtained by using complementary solid-state NMR methods. We demonstrate that the experimental conditions and procedures of sample crystallization determine the quality of solid-state NMR spectra and the internal mobility of the protein. Two-dimensional (2D) (13)C-(13)C and (15)N-(15)N correlation spectra reveal intra- and inter-residue dipolar connectivities and provide partial, site-specific assignments of (13)C and (15)N resonance signals. The secondary structure of the C-ter HsCen2/P17-XPC complex in a microcrystalline state appears similar to that found in solution. Conformational flexibility is probed through relaxation-compensated measurements of dipolar order parameters that exploit the dynamics of cross-polarization in multidimensional experiments. The extracted dipolar coupling constants and relevant order parameters reveal increased backbone flexibility of the loops except for residues involved in coordination with the Ca(2+) cation that stabilizes the hydrophobic pocket containing the peptide P17-XPC.
authors.affiliation: Ecole Normale Supérieure, Département de Chimie, 24 rue Lhomond, 75231 Paris, France.
authors.forename: Jose-Enrique
authors.initials: JE
authors.lastname: Herbert-Pucheta
authors.forename: Monique
authors.initials: M
authors.lastname: Chan-Huot
authors.forename: Luminita
authors.initials: L
authors.lastname: Duma
authors.forename: Daniel
authors.initials: D
authors.lastname: Abergel
authors.forename: Geoffrey
authors.initials: G
authors.lastname: Bodenhausen
authors.forename: Liliane
authors.initials: L
authors.lastname: Assairi
authors.forename: Yves
authors.initials: Y
authors.lastname: Blouquit
authors.forename: Jean-Baptiste
authors.initials: JB
authors.lastname: Charbonnier
authors.forename: Piotr
authors.initials: P
authors.lastname: Tekely
mesh_terms: D000595:Amino Acid Sequence; D002135:Calcium-Binding Proteins / Q000737:chemistry* / Q000378:metabolism*; D018797:Cell Cycle Proteins / Q000737:chemistry* / Q000378:metabolism*; D018360:Crystallography, X-Ray; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009068:Movement; D019906:Nuclear Magnetic Resonance, Biomolecular; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation
chemical_list: C472752:CETN2 protein, human; D002135:Calcium-Binding Proteins; D018797:Cell Cycle Proteins; D010455:Peptides