title: Cloning of glycoprotein D cDNA, which encodes the major subunit of the Duffy blood group system and the receptor for the Plasmodium vivax malaria parasite.
abstract: cDNA clones encoding the major subunit of the Duffy blood group were isolated from a human bone marrow cDNA library using a PCR-amplified DNA fragment encoding an internal peptide sequence of glycoprotein D (gpD) protein. The open reading frame of the 1267-bp cDNA clone indicated that gpD protein was composed of 338 amino acids, predicting a M(r) of 35,733, which was the same as a deglycosylated gpD protein. Portions of the predicted amino acid sequence, matched with six CNBr/pepsin peptides obtained from affinity-purified gpD protein. In ELISA analysis, an anti-Duffy murine monoclonal antibody reacted with a synthetic peptide deduced from the cDNA clone. Hydropathy analysis suggested the presence of 9 membrane-spanning alpha-helices. In bone marrow RNA blot analysis, the gpD cDNA detected a 1.27-kb mRNA in Duffy-positive but not in Duffy-negative individuals. It also identified the same size mRNA in adult kidney, adult spleen, and fetal liver; in brain, it detected a prominent 8.5-kb and a minor 2.2-kb mRNA. In Southern blot analysis, gpD cDNA identified a single gene in Duffy-positive and -negative individuals. Duffy-negative individuals, therefore, have the gpD gene, but it is not expressed in bone marrow. The same or a similar gene is active in adult kidney, adult spleen, and fetal liver of Duffy-positive individuals. Whether this is true in Duffy-negative individuals remains to be demonstrated. A GenBank sequence search yielded a significant protein sequence homology to human and rabbit interleukin-8 receptors.
authors.affiliation: Laboratory of Cell Biology, Lindsley F. Kimball Research Institute, New York, NY 10021.
authors.forename: A
authors.initials: A
authors.lastname: Chaudhuri
authors.forename: J
authors.initials: J
authors.lastname: Polyakova
authors.forename: V
authors.initials: V
authors.lastname: Zbrzezna
authors.forename: K
authors.initials: K
authors.lastname: Williams
authors.forename: S
authors.initials: S
authors.lastname: Gulati
authors.forename: A O
authors.initials: AO
authors.lastname: Pogo
mesh_terms: D000595:Amino Acid Sequence; D000953:Antigens, Protozoan; D001483:Base Sequence; D003001:Cloning, Molecular; D017931:DNA Primers / Q000737:chemistry; D018076:DNA, Complementary / Q000235:genetics; D004375:Duffy Blood-Group System / Q000235:genetics*; D015870:Gene Expression; D005796:Genes; D006801:Humans; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000235:genetics; D008969:Molecular Sequence Data; D015800:Protozoan Proteins; D012333:RNA, Messenger / Q000235:genetics; D011956:Receptors, Cell Surface / Q000235:genetics*; D015183:Restriction Mapping
chemical_list: D000953:Antigens, Protozoan; D017931:DNA Primers; D018076:DNA, Complementary; D004375:Duffy Blood-Group System; C066001:Duffy antigen binding protein, Plasmodium; D008562:Membrane Glycoproteins; D015800:Protozoan Proteins; D012333:RNA, Messenger; D011956:Receptors, Cell Surface