Aislamiento y caracterización bioquímica de la α-glucosidasa II del hongo patógeno Candida albicans Aislamiento y caracterización bioquímica de la α-glucosidasa II del hongo patógeno Candida albicans

• Main Authors: Arturo Flores Carreón, Alberto Flores-Martínez, Héctor Manuel Mora-Montes, Carlos Alberto Rangel Alfaro
• Format: Article
• Language: Spanish
• Published: Universidad de Guanajuato 2012-02-01
• Series: Acta Universitaria
• Subjects: Candida albicans; N-glicosilación; α-glucosidasa II; procesamiento proteolítico.
• Online Access: http://www.actauniversitaria.ugto.mx/index.php/acta/article/view/20

Alpha-glucosidase II participates in N-linked glycosylation of proteins. A soluble 47 kDa α-glucosidase II has been previously isolated from <em>C. albicans</em>; however, bioinformatics analysis indicate that native enzyme has a molecular mass of 100 kDa. In this study we assessed the effect of protease inhibitors on intracellular distribution of α-glucosidase II. Despite there was not a significant change in the enzyme distribution, α-glucosidase II activity was associated to a 83 or 47 kDa polypeptide in absence or presence of inhibitors, respectively. Soluble 83-kDa protein was purified by conventional methodology and its biochemical characteristics were similar to those reported for the 47 kDa enzyme. Thus, these results indicated the 83 kDa protein is an α-glucosidase II and also suggested it is a precursor of the 47 kDa enzyme previously reported.<br><span style="font-family: Times New Roman; font-size: small;"> </span><p class="MsoNormal" style="margin: 0cm 0cm 10pt;"><span style="color: black; line-height: 115%; font-size: 7.5pt; mso-bidi-font-family: "Bookman Old Style";"><span style="font-family: Calibri;">La </span></span><span style="color: black; line-height: 115%; font-family: "Times New Roman","serif"; font-size: 7.5pt;">α</span><span style="color: black; line-height: 115%; font-size: 7.5pt; mso-bidi-font-family: "Bookman Old Style";"><span style="font-family: Calibri;">-glucosidasa II participa en la ruta de la <em>N</em>-glicosilación de proteínas. En <em>Candida albi­cans </em>se ha aislado un polipéptido soluble de 47 kDa con actividad de </span></span><span style="color: black; line-height: 115%; font-family: "Times New Roman","serif"; font-size: 7.5pt;">α</span><span style="color: black; line-height: 115%; font-size: 7.5pt; mso-bidi-font-family: "Bookman Old Style";"><span style="font-family: Calibri;">-glucosidasa II; sin embargo, análisis bioinformáticos indican que la enzima nativa pudiera tener un peso mo­lecular de 100 kDa. En este trabajo se estudió el efecto de inhibidores de proteasas sobre la distribución intracelular de la α-glucosidasa II. Se demostró que la distribución intracelular no fue afectada significativamente, pero la actividad de la </span></span><span style="color: black; line-height: 115%; font-family: "Times New Roman","serif"; font-size: 7.5pt;">α</span><span style="color: black; line-height: 115%; font-size: 7.5pt; mso-bidi-font-family: "Bookman Old Style";"><span style="font-family: Calibri;">-glucosidasa II estuvo asociada a una proteína de 83 ó 47 kDa en ausencia o presencia de inhibidores de proteasas, res­pectivamente. La enzima soluble de 83 kDa se purificó por métodos convencionales y se demostró que presenta características bioquímicas similares a la enzima de 47 kDa. Estos datos confirmaron que la proteína de 83 kDa es una </span></span><span style="color: black; line-height: 115%; font-family: "Times New Roman","serif"; font-size: 7.5pt;">α</span><span style="color: black; line-height: 115%; font-size: 7.5pt; mso-bidi-font-family: "Bookman Old Style";"><span style="font-family: Calibri;">-glucosidasa II y sugieren que es precursora de la enzima de 47 kDa previamente descrita.</span></span></p><span style="font-family: Times New Roman; font-size: small;"> </span>