Molecular cloning, sequence, and expression of a human GDP-L-fucose:. beta. -D-galactoside 2-. alpha. -L-fucosyltransferase cDNA that can form the H blood group antigen

Journal Article · Sat Sep 01 00:00:00 EDT 1990 · Proceedings of the National Academy of Sciences of the United States of America; (United States)

DOI:https://doi.org/10.1073/pnas.87.17.6674· OSTI ID:5272391

Larsen, R D; Ernst, L K; Nair, R P; Lowe, J B ^[1]

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1. Univ. of Michigan, Ann Arbor (USA)

The authors have previously used a gene-transfer scheme to isolate a human genomic DNA fragment that determines expression of a GDP-L-fucose:{beta}D-galactoside 2-{alpha}-L-fucosyltransferase. Although this fragment determined expression of an {alpha}(1,2)FT whose kinetic properties mirror those of the human H blood group {alpha}(1,2)FT, their precise nature remained undefined. They describe here the molecular cloning, sequence, and expression of a human of cDNA corresponding to these human genomic sequences. When expressed in COS-1 cells, the cDNA directs expression of cell surface H structures and a cognate {alpha}(1,2)FT activity with properties analogous to the human H blood group {alpha}(1,2)FT. The cDNA sequence predicts a 365-amino acid polypeptide characteristic of a type II transmembrane glycoprotein with a domain structure analogous to that of other glycosyltransferases but without significant primary sequence similarity to these or other known proteins. To directly demonstrate that the cDNA encodes an {alpha}(1,2)FT, the COOH-terminal domain predicted to be Golgi-resident was expressed in COS-1 cells as a catalytically active, secreted, and soluble protein A fusion peptide. Southern blot analysis showed that this cDNA identified DNA sequences syntenic to the human H locus on chromosome 19. These results strongly suggest that this cloned {alpha}(1,2)FT cDNA represents the product of the human H blood group locus.

• OSTI ID: 5272391
• Journal Information: Proceedings of the National Academy of Sciences of the United States of America; (United States), Vol. 87:17; ISSN 0027-8424
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
GLYCOSYL TRANSFERASES
AMINO ACID SEQUENCE
DNA SEQUENCING
DNA-CLONING
GENE REGULATION
ANTIGENS
BLOOD GROUPS
GLYCOPROTEINS
HUMAN CHROMOSOME 19
MAN
PHOSPHORUS 32
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CHROMOSOMES
CLONING
DAYS LIVING RADIOISOTOPES
DNA HYBRIDIZATION
ENZYMES
HYBRIDIZATION
ISOTOPES
LIGHT NUCLEI
MAMMALS
MOLECULAR STRUCTURE
NUCLEI
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
PHOSPHORUS ISOTOPES
PRIMATES
PROTEINS
RADIOISOTOPES
STRUCTURAL CHEMICAL ANALYSIS
TRANSFERASES
VERTEBRATES
550201* - Biochemistry- Tracer Techniques