Glycoproteins with Gal alpha 4Gal are absent from human erythrocyte membranes, indicating that glycolipids are the sole carriers of blood group P activities.

Z Yang,J Bergström,K.A Karlsson

doi:10.1016/s0021-9258(17)36669-3

Z Yang, J Bergström + Show 1 more

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https://doi.org/10.1016/s0021-9258(17)36669-3

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Abstract

The antigenic determinants of the blood group P family (P1, P, Pk, and LKE antigens) are chemically based on Gal alpha 4Gal. For human erythrocytes it has been claimed that the majority of P1 determinants are expressed in glycoproteins, mainly band 4.5 (Haselberger, C. G., and Schenkel-Brunner, H. (1982) FEBS Lett. 149, 126-128). In the present work, the existence of Gal alpha 4Gal in glycoproteins of erythrocyte membranes (ghosts) of P1 positive and negative human individuals was carefully analyzed on replicas of sodium dodecyl sulfate-polyacrylamide electrophoresis gels using specific reagents (Escherichia coli HB101/pDC1 expressing the Pap gene and monoclonal antibodies with specificities for P1 and Pk antigens). No binding to glycoproteins was detected with any of these ligands when the ghosts had been pretreated with butanol to remove glycolipids. Therefore, all antigenic determinants of the P blood group family on human red cells are exclusively expressed in glycolipids and are absent from glycoproteins.

Highlights

Chemical spacers used for the synthesis of for P, and Pkantigens).No binding to glycoproteins wasneoglycoproteins(BioCarb)were: PAP, p-aminophenyl; CETE, 2-(2-cardetected with anyof these ligands when the ghosts hadbomethoxyethy1thio)ethyl; and AF'D, acetylphenylenediamine
All antigenic determinants of the P blood groupfamilyonhumanred cells areexclusivelyexpressed in glycolipids and are absent from glycoproteins
Ceramide glycanasefrom leech(Macrobdelladecora) was purchased from Boehringer Mannheim (Mannheim,Germany).0.5 microunit was added to the ch1oroform:methanol extraction product, both a terminal and an internal Gala4Gal sequence

Summary

Introduction

Butanol Deatment of the ErythrocyteGhosts-To remove glycolipids containing 200 mM NaCI, 0.05%Tween 20(pH 8.0)).The second labeled from erythrocyte membranes, the ghosts prepared as described above anti-mouse antibody, with a final concentration of 1-2 pg/ml and radiowere treated by 1-butanol. The supernatant was treated by butanol as described above to ensure the complete removalof glycolipids, and the resulting pellet was dissolved in Tris-HC1 buffercontaining SDS and 2-merglycolipids addedtothe nitrocellulose membrane were all bound by E.coli.

Results

Conclusion