Miyagi T, Tsuiki S
Biochim Biophys Acta. 1981 Sep 15;661(1):148-57. doi: 10.1016/0005-2744(81)90094-2.
When homogenates of rat liver and hepatomas were centrifuged at 78 000 X g, over 90% of liver N-acetylglucosaminyltransferase assayed with beta-galactosidase- and beta-N-acetylhexosaminidase-treated asialofetuin as acceptor was recovered in the particulate fraction, while as much as 24% of hepatoma transferase was in the supernatant fraction. The particulate transferase solubilized by 0.2% sodium deoxycholate emerged from a DEAE-cellulose column at 0.04 M NaCl (transferase A). The supernatant fractions from all the hepatomas tested contained a second N-acetylglucosaminyltransferase eluted from the column at 0.02 M NaCl (transferase B). Transferase B was absent from liver supernatant fraction. The activities of these transferases toward various acceptors and the effect of beta-N-acetylhexosaminidase on their products suggest that both transferases are UDP-N-acetylglucosamine : alpha-mannoside beta-N-acetylglucosaminyltransferase. Although ovalbumin and glycopeptide V, which was isolated from pronase digest of ovalbumin, were good acceptors, transferase A utilized ovalbumin and glycopeptide V with apparent Km values of 0.44 and 0.33 mM, respectively, whereas the corresponding values for transferase B were 4.5 and 0.050 mM.
当大鼠肝脏和肝癌匀浆在78000×g下离心时，以经β-半乳糖苷酶和β-N-乙酰己糖胺酶处理的去唾液酸胎球蛋白作为受体测定的肝脏N-乙酰葡糖胺转移酶，超过90%回收在颗粒部分，而肝癌转移酶高达24%在上清部分。用0.2%脱氧胆酸钠溶解的颗粒转移酶在0.04M NaCl时从DEAE-纤维素柱洗脱（转移酶A）。所有测试肝癌的上清部分含有第二种N-乙酰葡糖胺转移酶，在0.02M NaCl时从柱上洗脱（转移酶B）。肝脏上清部分没有转移酶B。这些转移酶对各种受体的活性以及β-N-乙酰己糖胺酶对其产物的影响表明，两种转移酶都是UDP-N-乙酰葡糖胺：α-甘露糖苷β-N-乙酰葡糖胺转移酶。虽然卵清蛋白和从卵清蛋白的链霉蛋白酶消化物中分离的糖肽V是良好的受体，但转移酶A利用卵清蛋白和糖肽V时的表观Km值分别为0.44和0.33mM，而转移酶B的相应值为4.5和0.050mM。
