Nishida Chiyomi, Nishiyama Chiharu, Satoh Koichi, Hara Mutsuko, Itoh Yukio, Ogawa Hideoki, Okumura Ko
Department of Forensic Medicine, Juntendo University School of Medicine, Tokyo, Japan.
Leg Med (Tokyo). 2010 Jul;12(4):172-6. doi: 10.1016/j.legalmed.2010.03.005. Epub 2010 May 14.
A/B-transferase is a glycosyltransferase that transfers a sugar substrate onto H-antigen resulting in the synthesis of glycoproteins and glycolipids termed A/B-antigens. The ABO blood group (ABO) gene encoding A/B-transferase possesses numerous polymorphisms affecting the specificity and/or activity of the enzyme. The relationship between genotype and phenotype is very complicated, except for those of some critical polymorphisms. In order to establish a system for evaluating the effect of each polymorphism on the transferase function, an A- or B-transferase cDNA expressing vector was introduced into HeLa cells, a cell line that do not possess endogenous A/B-transferase activity. We successfully detected substrate-specific transferase activity in the cells and in the culture medium. Furthermore, in three different assays, each corresponding A- or B-antigen was detected in the transfectants with high sensitivity. Accordingly, the present study demonstrates a possibility that A/B-transferase variants may be characterized by using this method.
A/B-转移酶是一种糖基转移酶,它将糖底物转移到H抗原上,从而合成称为A/B抗原的糖蛋白和糖脂。编码A/B-转移酶的ABO血型(ABO)基因具有许多影响该酶特异性和/或活性的多态性。除了一些关键多态性的情况外,基因型和表型之间的关系非常复杂。为了建立一个评估每种多态性对转移酶功能影响的系统,将A或B转移酶cDNA表达载体导入HeLa细胞,这是一种不具有内源性A/B转移酶活性的细胞系。我们成功地在细胞和培养基中检测到了底物特异性转移酶活性。此外,在三种不同的检测方法中,在转染子中以高灵敏度检测到了各自相应的A或B抗原。因此,本研究证明了使用这种方法表征A/B转移酶变体的可能性。