Metzner S I, Sousa M C, Hellman U, Cazzulo J J, Parodi A J
Instituto de Investigaciones Bioquímicas Fundación Campomar, Buenos Aires, Argentina.
Cell Mol Biol (Noisy-le-grand). 1996 Jul;42(5):631-5.
A simple method for specifically radiolabeling high mannose-type oligosaccharides linked to protein backbones has been developed. The method is based on the fact that incubation of rat liver UDP-Glc:glycoprotein glucosyltransferase, glucose-labelled UDP-Glc and a denatured high mannose-type glycoprotein target leads to the glucosylation of the oligosaccharide. In the case described here it allowed to follow easily the purification, by HPLC and affinity chromatography, of labelled glycopeptides obtained by controlled proteolysis of cruzipain, a cysteine proteinase isolated from the human pathogen Trypanosoma cruzi. It was thus determined that the N-glycosylation site in Asn33 of cruzipain is occupied by high mannose-type oligosaccharides.
已经开发出一种特异性放射性标记与蛋白质骨架相连的高甘露糖型寡糖的简单方法。该方法基于这样一个事实，即大鼠肝脏UDP-葡萄糖：糖蛋白葡糖基转移酶、葡萄糖标记的UDP-葡萄糖与变性的高甘露糖型糖蛋白靶标一起孵育会导致寡糖的糖基化。在本文所述的情况下，通过高效液相色谱法（HPLC）和亲和色谱法，可以轻松跟踪通过对从人类病原体克氏锥虫分离的半胱氨酸蛋白酶克氏锥虫蛋白酶进行可控蛋白水解而获得的标记糖肽的纯化过程。由此确定，克氏锥虫蛋白酶Asn33处的N-糖基化位点被高甘露糖型寡糖占据。
