TY - JOUR
T1 - Probing structural and motional features of the c-terminal part of the human Centrin 2/P17-XPC microcrystalline complex by solid-state NMR spectroscopy
AU - Herbert-Pucheta, Jose Enrique
AU - Chan-Huot, Monique
AU - Duma, Luminita
AU - Abergel, Daniel
AU - Bodenhausen, Geoffrey
AU - Assairi, Liliane
AU - Blouquit, Yves
AU - Charbonnier, Jean Baptiste
AU - Tekely, Piotr
PY - 2012/12/20
Y1 - 2012/12/20
N2 - 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) 13C-13C and 15N- 15N correlation spectra reveal intra-and inter-residue dipolar connectivities and provide partial, site-specific assignments of 13C and 15N 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 Ca2+ cation that stabilizes the hydrophobic pocket containing the peptide P17-XPC.
AB - 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) 13C-13C and 15N- 15N correlation spectra reveal intra-and inter-residue dipolar connectivities and provide partial, site-specific assignments of 13C and 15N 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 Ca2+ cation that stabilizes the hydrophobic pocket containing the peptide P17-XPC.
UR - http://www.scopus.com/inward/record.url?scp=84871598384&partnerID=8YFLogxK
U2 - 10.1021/jp3099472
DO - 10.1021/jp3099472
M3 - Artículo
C2 - 23190348
AN - SCOPUS:84871598384
SN - 1520-6106
VL - 116
SP - 14581
EP - 14591
JO - Journal of Physical Chemistry B
JF - Journal of Physical Chemistry B
IS - 50
ER -