Establishment of a simple detection system for blood group ABO-specific transferase activity in DNA-transfected cells.
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.
Nishida
Chiyomi
C
Department of Forensic Medicine, Juntendo University School of Medicine, Tokyo, Japan.
Nishiyama
Chiharu
C
Satoh
Koichi
K
Hara
Mutsuko
M
Itoh
Yukio
Y
Ogawa
Hideoki
H
Okumura
Ko
K
D000017:ABO Blood-Group System / Q000235:genetics*; D001788:Blood Grouping and Crossmatching / Q000379:methods*; D005434:Flow Cytometry; D005822:Genetic Vectors; D006367:HeLa Cells / Q000201:enzymology; D006801:Humans; D007564:Japan; D014162:Transfection; D014166:Transferases / Q000032:analysis / Q000235:genetics*
D000017:ABO Blood-Group System; D014166:Transferases