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Originally published In Press as doi:10.1074/jbc.M000728200 on March 21, 2000

J. Biol. Chem., Vol. 275, Issue 22, 16723-16729, June 2, 2000
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Cloning and Expression of the Histo-blood Group Pk UDP-galactose:Galbeta 1-4Glcbeta 1-Cer alpha 1,4-Galactosyltransferase
MOLECULAR GENETIC BASIS OF THE p PHENOTYPE*

Rudi SteffensenDagger §, Karine Carlier, Joelle Wiels, Steven B. Levery||, Mark Stroud**, Bertil CedergrenDagger Dagger , Birgitta Nilsson SojkaDagger Dagger , Eric P. BennettDagger , Casper Jersild§, and Henrik ClausenDagger §§

From the Dagger  School of Dentistry, University of Copenhagen, Nørre Allé 20, 2200 Copenhagen N, Denmark, the § Regional Center for Blood Transfusion and Clinical Immunology, Aalborg Hospital, 9000 Aalborg, Denmark,  CNRS UMR 1598, Institut Gustave Roussy, 94805 Villejuif Cedex, France, the || Complex Carbohydrate Research Center, University of Georgia, Athens, Georgia 30602, the ** Department of Cell Surface Biochemistry, Northwest Hospital, Seattle, Washington 98125, and the Dagger Dagger  Department of Transfusion Medicine, Umeå University Hospital, S-901 85 Umeå, Sweden

The molecular genetic basis of the P histo-blood group system has eluded characterization despite extensive studies of the biosynthesis of the P1, P, and Pk glycolipids. The main controversy has been whether a single or two distinct UDP-Gal:Galbeta 1-R 4-alpha -galactosyltransferases catalyze the syntheses of the structurally related P1 and Pk antigens. The P1 polymorphism is linked to 22q11.3-ter. Data base searches with the coding region of an alpha 4GlcNAc-transferase identified a novel homologous gene at 22q13.2 designated alpha 4Gal-T1. Expression of full coding constructs of alpha 4Gal-T1 in insect cells revealed it encoded Pk but not P1 synthase activity. Northern analysis showed expression of the transcript correlating with Pk synthase activity and antigen expression in human B cell lines. Transfection of Pk-negative Namalwa cells with alpha 4Gal-T1 resulted in strong Pk expression. A single homozygous missense mutation, M183K, was found in six Swedish individuals of the rare p phenotype, confirming that alpha 4Gal-T1 represented the Pk gene. Sequence analysis of the coding region of alpha 4Gal-T1 in P1+/- individuals did not reveal polymorphisms correlating with P1P2 typing.

* This work was supported by the PhD Foundation Aalborg Sygehus, Aalborg Frivillige Bloddonorers Foundation, Nordjyllands Amts Foundation, Det Obelse Familiefond, Peder Kristen Tøfting, and Dagmar Tøftings Foundation, Ebba and Aksel Schølins Foundation, the Danish Cancer Society, the Velux Foundation, the Danish Research Council, Grant 5 P41 RR05351 from the National Institutes of Health Resource Center for Biomedical Complex Carbohydrates, and Grant ARC9588 from the Association pour la Recherche sur le Cancer.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EMBL Data Bank with accession number(s) AJ245581.

§§ To whom correspondence should be addressed. Tel.: 45-35326835; Fax: 45-35326505; E-mail: henrik.clausen@odont.ku.dk.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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