Tabatabai L B
National Animal Disease Center, USDA, ARS, Ames, Iowa 50010, USA.
Biochem Biophys Res Commun. 1995 Jul 26;212(3):981-7. doi: 10.1006/bbrc.1995.2066.
A soluble bovine glycoprotein, fetuin, was used as an alternative substrate to identify O-sialoglycoprotease activity in culture supernatant protein fractions of Pasteurella haemolytica. An aliquot of a 24-hour incubation mixture containing fetuin and O-sialoglycoprotease was denatured and examined after gradient sodium dodecyl polyacrylamide gel electrophoresis. The Coomassie-Brilliant-Blue-stained gel was examined for the disappearance of the fetuin band at an apparent molecular mass of 64 KDa. Four major hydrolysis products were identified: an N-terminal fragment of 45 kDa, a 20 kDa fragment at Val50, and two C-terminal fragments at Val273 and His287.
一种可溶性牛糖蛋白胎球蛋白被用作替代底物,以鉴定溶血巴斯德氏菌培养上清蛋白组分中的O-唾液酸糖蛋白酶活性。含有胎球蛋白和O-唾液酸糖蛋白酶的24小时孵育混合物的一份等分试样经变性处理后,在梯度十二烷基硫酸钠聚丙烯酰胺凝胶电泳后进行检测。检查考马斯亮蓝染色的凝胶,看表观分子量为64 kDa的胎球蛋白条带是否消失。鉴定出四种主要水解产物:一个45 kDa的N端片段、一个位于Val50的20 kDa片段以及位于Val273和His287的两个C端片段。