INVESTIGADORES
CHEHIN Rosana Nieves
artículos
Título:
Maintenance and thermal stabilization of NADH dehydrogenase-2 conformation upon elimination of its C-terminal region
Autor/es:
VILLEGAS JM, TORRES-BUGEAU CM, CHEHÍN R, BURGOS MI, FIDELIO GD, RINTOUL MR, RAPISARDA VA
Revista:
BIOCHIMIE
Editorial:
ELSEVIER FRANCE-EDITIONS SCIENTIFIQUES MEDICALES ELSEVIER
Referencias:
Lugar: Paris; Año: 2013 vol. 95 p. 382 - 387
ISSN:
0300-9084
Resumen:
Development of an artificial enzyme with activity and structure comparable to that of natural enzymes is an important goal in biological chemistry. Respiratory NADH dehydrogenase-2 (NDH-2) of Escherichia coli is a peripheral membrane-bound flavoprotein, belonging to a group of enzymes with scarce structural information. By eliminating the C-terminal region of NDH-2, a water soluble version with significant enzymatic activity was previously obtained. Here, NDH-2 structural features were established, in comparison to those of the truncated version. Far-UV circular dichroism, Fourier transform infrared spectroscopy and limited proteolysis analysis showed that the overall structure of both proteins was similar at 30 °C. Experimental data agree with the predicted NDH-2 structure (PDB: 1OZK). The absence of C-terminal region stabilized in ∼5-10 °C the truncated protein conformation. However, truncation impaired enzymatic activity at low temperatures, probably due to the weak interaction of the mutant protein with FAD cofactor.