Lee K N, Birckbichler P J, Patterson M K
Biomedical Division, Samuel Roberts Noble Foundation, Inc., Ardmore, Oklahoma 73402.
Biochem Biophys Res Commun. 1989 Aug 15;162(3):1370-5. doi: 10.1016/0006-291x(89)90825-5.
Homogeneous guinea pig liver transglutaminase was purified from a commercially available enzyme preparation by affinity chromatography on GTP-agarose. The purified transglutaminase exhibited a single band of apparent Mr = 80,000 on sodium dodecyl sulfate polyacrylamide gel and Western blotting and had enzyme activity of both transglutaminase and GTPase. The guinea pig liver transglutaminase has an apparent Km value of 4.4 microM for GTPase activity. GTPase activity was inhibited by guanine nucleotides in order GTP-gamma-S greater than GDP, but not by GMP. These results demonstrate that purified guinea pig liver transglutaminase catalyzes GTP hydrolysis.
通过在GTP-琼脂糖上进行亲和层析，从市售酶制剂中纯化出均一的豚鼠肝脏转谷氨酰胺酶。纯化的转谷氨酰胺酶在十二烷基硫酸钠聚丙烯酰胺凝胶和蛋白质免疫印迹上呈现出一条表观分子量为80,000的条带，并且具有转谷氨酰胺酶和GTP酶的活性。豚鼠肝脏转谷氨酰胺酶的GTP酶活性的表观Km值为4.4微摩尔。GTP酶活性被鸟嘌呤核苷酸按GTP-γ-S＞GDP的顺序抑制，但不被GMP抑制。这些结果表明纯化的豚鼠肝脏转谷氨酰胺酶催化GTP水解。
