TY - JOUR
T1 - Crystal structure and molecular dynamics studies of human purine nucleoside phosphorylase complexed with 7-deazaguanine
AU - Caceres, Rafael Andrade
AU - Timmers, Luis Fernando Saraiva Macedo
AU - Pauli, Ivani
AU - Gava, Lisandra Marques
AU - Ducati, Rodrigo Gay
AU - Basso, Luiz Augusto
AU - Santos, Diógenes Santiago
AU - de Azevedo, Walter Filgueira
N1 - Funding Information:
This work was supported by the National Institute of Science and Technology on Tuberculosis (Decit/SCTIE/MS-MCT-CNPq-FNDCT-CAPES) and Millennium Initiative Program (CNPq), Brazil . W.F.A. (300851/98-7), D.S.S. (304051/1975-06), and L.A.B. (520182/99-5) are research career awardees of the National Council for Scientific and Technological Development of Brazil (CNPq). R.A.C., L.F.S.M.T., and I.P. would like to thank CNPq for the fellowship.
PY - 2010/3
Y1 - 2010/3
N2 - In humans, purine nucleoside phosphorylase (HsPNP) is responsible for degradation of deoxyguanosine, and genetic deficiency of this enzyme leads to profound T-cell mediated immunosuppression. HsPNP is a target for inhibitor development aiming at T-cell immune response modulation. Here we report the crystal structure of HsPNP in complex with 7-deazaguanine (HsPNP:7DG) at 2.75 Å. Molecular dynamics simulations were employed to assess the structural features of HsPNP in both free form and in complex with 7DG. Our results show that some regions, responsible for entrance and exit of substrate, present a conformational variability, which is dissected by dynamics simulation analysis. Enzymatic assays were also carried out and revealed that 7-deazaguanine presents a lower inhibitory activity against HsPNP (Ki = 200 μM). The present structure may be employed in both structure-based design of PNP inhibitors and in development of specific empirical scoring functions.
AB - In humans, purine nucleoside phosphorylase (HsPNP) is responsible for degradation of deoxyguanosine, and genetic deficiency of this enzyme leads to profound T-cell mediated immunosuppression. HsPNP is a target for inhibitor development aiming at T-cell immune response modulation. Here we report the crystal structure of HsPNP in complex with 7-deazaguanine (HsPNP:7DG) at 2.75 Å. Molecular dynamics simulations were employed to assess the structural features of HsPNP in both free form and in complex with 7DG. Our results show that some regions, responsible for entrance and exit of substrate, present a conformational variability, which is dissected by dynamics simulation analysis. Enzymatic assays were also carried out and revealed that 7-deazaguanine presents a lower inhibitory activity against HsPNP (Ki = 200 μM). The present structure may be employed in both structure-based design of PNP inhibitors and in development of specific empirical scoring functions.
KW - 7-Deazaguanine
KW - Enzymatic assay
KW - Molecular dynamics
KW - Purine nucleoside phosphorylase
KW - Virtual screening
KW - X-ray diffraction
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U2 - 10.1016/j.jsb.2009.11.010
DO - 10.1016/j.jsb.2009.11.010
M3 - Article
C2 - 19932753
AN - SCOPUS:77049096015
SN - 1047-8477
VL - 169
SP - 379
EP - 388
JO - Journal of Structural Biology
JF - Journal of Structural Biology
IS - 3
ER -