Human thrombomodulin: complete cDNA sequence and chromosome localization of the gene
A human umbilical vein endothelial cell cDNA library in lambdagt11 was screened for expression of thrombomodulin antigens with affinity-purified rabbit polyclonal anti-thrombomodulin immunoglobulin G (IgG) and mouse monoclonal anti-human thrombomodulin IgG. Among 7 million recombinant clones screened, 12 were recognized by both antibodies. Two of these, lambdaHTm10 and lambdaHTm12, were shown to encode thrombomodulin by comparison of the amino acid sequence deduced from the nucleotide sequence to the amino acid sequence determined directly from tryptic peptides of thrombomodulin. Thrombomodulin mRNA was estimated to be 3.7 kilobases in length by Northern blot analysis of endothelial cell and placental poly(A) + RNA. Thrombomodulin mRNA was not detected in human brain, HepG2 hepatoma cells, or the monocytic U937 cell line. Additional cDNA clones were selected by hybridization with the 1.2-kilobase insert of lambdaHTm10. One isolate, lambdaHTm15, contained a 3693 base pair cDNA insert with an apparent 5'-noncoding region of 146 base pairs, an open reading frame of 1725 base pairs, a stop codon, a 3'-noncoding region of 1779 base pairs, and a poly(A) tail of 40 base pairs. The cDNA sequence encodes a 60.3-kDa protein of 575 amino acids. The organization of thrombomodulin is similar to that of the low-density lipoprotein receptor, and the protein is homologous to a large number of other proteins that also contain EGF-like domains, including factor VII, factor IX, factor X, factor XII, protein C, tissue plasminogen activator, and urokinase. The gene for thrombomodulin has been localized to chromosome 20 by hybridization of cDNA probes to purified human chromosomes.
- Research Organization:
- Washington Univ. School of Medicine, St. Louis, MO
- OSTI ID:
- 6080218
- Journal Information:
- Biochemistry; (United States), Vol. 26:14
- Country of Publication:
- United States
- Language:
- English
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DNA SEQUENCING
THROMBIN
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GENETIC MAPPING
LABELLED COMPOUNDS
MAN
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SULFUR 35
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DRUGS
ENZYMES
EVEN-ODD NUCLEI
HEMATOLOGIC AGENTS
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MAMMALS
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MEMBRANE PROTEINS
MOLECULAR STRUCTURE
NUCLEI
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ORGANIC COMPOUNDS
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550201* - Biochemistry- Tracer Techniques
550401 - Genetics- Tracer Techniques