Characterization of the histo-blood group O-2 gene and its protein product

Background and Objectives: This study aimed to show the full sequence and f unction of the O-2 allele, and investigate whether it accounts for the inco mpatible expression of A antigens in gastric carcinomas of blood group O pe rsons. Materials and Methods: By PCR, we determined the ABO genotype of gro up O subjects (76 gastric carcinoma patients and 165 blood donors). Two exp ression constructs, encoding either the putative soluble or full-length O-2 protein, were used to transfect Sf9 cells. The expression and the activity of the O-2 protein were analysed by immunohistochemistry and enzymatic ass ays, respectively. Results: No significant difference was detectable betwee n the O-2 allele frequency in gastric carcinoma patients (3.9%) and blood d onors (4.2%). Sequencing analysis of the O-2 allele revealed an intact read ing frame identical to that of A transferase except for four nucleotide sub stitutions. O-2-transfected Sf9 cells and gastric carcinomas genotyped as ( OO2)-O-1 both expressed a protein recognized by anti-A/B transferase monocl onal antibodies. In enzymatic assays, the O-2 protein failed to show measur able A transferase activity. Conclusion: The O-2 allele has an intact readi ng frame encoding a protein immunologically related to A/B transferases and enzymatically inactive. Further, our data gave no indication that the O-2 allele is related to the phenomenon of incompatible A antigen expression in gastric cancer. Copyright (C) 2000 S. Karger AG, Basel.