The P1 histo-blood group antigen is present on human red blood cell glycoproteins.

BACKGROUND The P1 antigen was first described in 1927 and belongs to the P1PK histo-blood group system, together with P-k and NOR. The A4GALT-encoded 4-alpha-galactosyltransferase synthesizes these antigens and has been considered to extend glycolipids exclusively. However, contradicting studies have been published regarding the presence of P1 on human glycoproteins. In other species, P1 occurs on glycoproteins. Furthermore, human ABH antigens occur on both glycolipids and glycoproteins and are biochemically related to P1. Thus, we hypothesized that P1 is present on RBC glycoproteins in humans. STUDY DESIGN AND METHODS RBCs of known P-1/P-2 status (phenotype and rs8138197 genotype) were used. The RBC surface glycans were modified with alpha-galactosidases, papain, and/or peptide-N-glycosidase F. RBC membrane proteins were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis/immunoblot. A new P-1/P-2-allelic discrimination assay based on rs5751348 was validated. RESULTS P1 occurs on various glycoproteins, seen as smearlike patterns in anti-P1-stained immunoblots with RBC membranes of P-1 but not P-2 or p phenotype. There was a significant difference between the staining of P-1-homozygous and P-1-heterozygous RBCs ((PP1)-P-1 > (PP2)-P-1), as well as intragenotypic variation. Immunoblotting banding patterns show major carriers at approximately 50 and 100 kDa. P1 staining was lost after treatment of RBCs with alpha-galactosidase of broad Gal alpha-1,3/4/6-specificity. Peptide-N-glycosidase F treatment reduced the P1 signal, while papain or alpha-1,3-specific galactosidase did not. P-1/P-2 status was confirmed by a new rs5751348 assay. CONCLUSION Our data indicate that the P1 antigen can reside on human RBC glycoproteins. Glycosidase studies suggest that at least part of the epitopes occur on N-glycans.