TY - JOUR
T1 - Identification of the NADP+ structural binding site and coenzyme effect on the fused G6PD::6PGL protein from giardia lamblia
AU - Morales-Luna, Laura
AU - González-Valdez, Abigail
AU - Sixto-López, Yudibeth
AU - Correa-Basurto, José
AU - Hernández-Ochoa, Beatriz
AU - Cárdenas-Rodríguez, Noemí
AU - Castillo-Rodríguez, Rosa Angélica
AU - Ortega-Cuellar, Daniel
AU - Arreguin-Espinosa, Roberto
AU - de la Cruz, Verónica Pérez
AU - Serrano-Posada, Hugo
AU - Centeno-Leija, Sara
AU - Rocha-Ramírez, Luz María
AU - Sierra-Palacios, Edgar
AU - Montiel-González, Alba Mónica
AU - Rufino-González, Yadira
AU - Marcial-Quino, Jaime
AU - Gómez-Manzo, Saúl
N1 - Publisher Copyright:
© 2019 by the authors. Licensee MDPI, Basel, Switzerland.
PY - 2020/1
Y1 - 2020/1
N2 - Giardia lambia is a flagellated protozoan parasite that lives in the small intestine and is the causal agent of giardiasis. It has been reported that G. lamblia exhibits glucose-6-phosphate dehydrogenase (G6PD), the first enzyme in the pentose phosphate pathway (PPP). Our group work demonstrated that the g6pd and 6pgl genes are present in the open frame that gives rise to the fused G6PD::6PGL protein; where the G6PD region is similar to the 3D structure of G6PD in Homo sapiens. The objective of the present work was to show the presence of the structural NADP+ binding site on the fused G6PD::6PGL protein and evaluate the effect of the NADP+ molecule on protein stability using biochemical and computational analysis. A protective effect was observed on the thermal inactivation, thermal stability, and trypsin digestions assays when the protein was incubated with NADP+. By molecular docking, we determined the possible structural-NADP+ binding site, which is located between the Rossmann fold of G6PD and 6PGL. Finally, molecular dynamic (MD) simulation was used to test the stability of this complex; it was determined that the presence of both NADP+ structural and cofactor increased the stability of the enzyme, which is in agreement with our experimental results.
AB - Giardia lambia is a flagellated protozoan parasite that lives in the small intestine and is the causal agent of giardiasis. It has been reported that G. lamblia exhibits glucose-6-phosphate dehydrogenase (G6PD), the first enzyme in the pentose phosphate pathway (PPP). Our group work demonstrated that the g6pd and 6pgl genes are present in the open frame that gives rise to the fused G6PD::6PGL protein; where the G6PD region is similar to the 3D structure of G6PD in Homo sapiens. The objective of the present work was to show the presence of the structural NADP+ binding site on the fused G6PD::6PGL protein and evaluate the effect of the NADP+ molecule on protein stability using biochemical and computational analysis. A protective effect was observed on the thermal inactivation, thermal stability, and trypsin digestions assays when the protein was incubated with NADP+. By molecular docking, we determined the possible structural-NADP+ binding site, which is located between the Rossmann fold of G6PD and 6PGL. Finally, molecular dynamic (MD) simulation was used to test the stability of this complex; it was determined that the presence of both NADP+ structural and cofactor increased the stability of the enzyme, which is in agreement with our experimental results.
KW - Docking
KW - G6PD
KW - Giardia lamblia
KW - NADP structural binding site
KW - Stability
UR - http://www.scopus.com/inward/record.url?scp=85077327369&partnerID=8YFLogxK
U2 - 10.3390/biom10010046
DO - 10.3390/biom10010046
M3 - Artículo
C2 - 31892224
SN - 2218-273X
VL - 10
JO - Biomolecules
JF - Biomolecules
IS - 1
M1 - 46
ER -