A new high-molecular-weight glycophorin C variant with a duplication of exon 2 in the glycophorin C gene.

Abstract

Glycophorin C (GPC) and glycophorin D (GPD) are closely related sialoglycoproteins in the human red blood cell (RBC) membrane. Both are thought to be encoded by the GPC gene (GYPC). We report here the new GPC variant, MAT, with a high-molecular-weight form of GPC and GPD. The murine monoclonal antibody to GPC (CBC-96), which had specificity for the N-terminal region of GPC, gave a stronger reactivity with the MAT RBCs than did normal RBCs in direct agglutination tests. Immunoblotting of the MAT RBC membranes with anti-GPC antibodies showed the apparent molecular weight of GPC.MAT and GPD.MAT was 5000 greater than that of their normal counterparts. cDNA was synthesized from total RNA obtained from three unrelated, heterozygous MAT blood donors and analysed by the polymerase chain reaction with primers that spanned sequences encoded by GYPC. Two fragments were generated: one was 510 bp, the other was 453 bp and corresponded to the normal GPC. Sequencing of the mutant 510-bp fragment showed an insert of 57 nucleotides that corresponds to the entire sequence of exon 2 in GYPC. These results show that MAT is the result of a duplication of exon 2 in GYPC, which probably encodes the two high-molecular weight forms GPC.MAT and GPD.MAT. The MAT mutation is found with a frequency of 0.02% in the Japanese population.