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Dr(a-) polymorphism of decay accelerating factor. Biochemical, functional, and molecular characterization and production of allele-specific transfectants.

Publication ,  Journal Article
Lublin, DM; Thompson, ES; Green, AM; Levene, C; Telen, MJ
Published in: J Clin Invest
June 1991

The Dra antigen belongs to the Cromer-related blood group system, a series of antigens on decay accelerating factor (DAF), a glycosyl-phosphatidylinositol-anchored membrane protein that protects host cells from complement-mediated damage. We studied the rare inherited Dr(a-) phenotype to ascertain the associated biochemical and functional changes in DAF and to characterize the basis for this polymorphism. Radioimmunoassay assay and flow cytometric analysis of Dr(a-) erythrocytes demonstrated 40% of normal surface expression of DAF but normal levels of several other glycosyl-phosphatidylinositol-anchored proteins, distinguishing this phenotype from that of paroxysmal nocturnal hemoglobinuria. Western blots confirmed this reduced DAF expression and indicated a slightly faster mobility of the molecule on SDS-PAGE. Despite the reduced DAF expression, Dr(a-) erythrocytes functioned normally in the complement lysis sensitivity assay. Utilization of the polymerase chain reaction to amplify mononuclear cell genomic DNA from three unrelated Dr(a-) individuals demonstrated that a point mutation underlies the Dr(a-) phenotype: a C to T change in nucleotide 649 resulting in a serine165 to leucine change. This defines the Drb allele of DAF, which can be distinguished from Dra by a Taq I restriction fragment length polymorphism. We created transfected Chinese hamster ovary cell lines expressing either the Dra or the Drb allelic form of DAF. These allele-specific transfectants were tested by inhibition of hemagglutination or flow cytometry and confirmed the specificity of anti-Dra alloantisera. The allele-specific transfectants could form the basis of a new serological approach to immunohematology.

Duke Scholars

Published In

J Clin Invest

DOI

ISSN

0021-9738

Publication Date

June 1991

Volume

87

Issue

6

Start / End Page

1945 / 1952

Location

United States

Related Subject Headings

  • Transfection
  • Polymorphism, Restriction Fragment Length
  • Polymorphism, Genetic
  • Polymerase Chain Reaction
  • Phosphatidylinositols
  • Oligonucleotides
  • Molecular Sequence Data
  • Membrane Proteins
  • Immunology
  • Humans
 

Citation

APA
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ICMJE
MLA
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Lublin, D. M., Thompson, E. S., Green, A. M., Levene, C., & Telen, M. J. (1991). Dr(a-) polymorphism of decay accelerating factor. Biochemical, functional, and molecular characterization and production of allele-specific transfectants. J Clin Invest, 87(6), 1945–1952. https://doi.org/10.1172/JCI115220
Lublin, D. M., E. S. Thompson, A. M. Green, C. Levene, and M. J. Telen. “Dr(a-) polymorphism of decay accelerating factor. Biochemical, functional, and molecular characterization and production of allele-specific transfectants.J Clin Invest 87, no. 6 (June 1991): 1945–52. https://doi.org/10.1172/JCI115220.
Lublin, D. M., et al. “Dr(a-) polymorphism of decay accelerating factor. Biochemical, functional, and molecular characterization and production of allele-specific transfectants.J Clin Invest, vol. 87, no. 6, June 1991, pp. 1945–52. Pubmed, doi:10.1172/JCI115220.

Published In

J Clin Invest

DOI

ISSN

0021-9738

Publication Date

June 1991

Volume

87

Issue

6

Start / End Page

1945 / 1952

Location

United States

Related Subject Headings

  • Transfection
  • Polymorphism, Restriction Fragment Length
  • Polymorphism, Genetic
  • Polymerase Chain Reaction
  • Phosphatidylinositols
  • Oligonucleotides
  • Molecular Sequence Data
  • Membrane Proteins
  • Immunology
  • Humans