title: Identification and characterization of a thrombomodulin gene mutation coding for an elongated protein with reduced expression in a kindred with myocardial infarction.
abstract: Thrombomodulin is an endothelial cell receptor for thrombin. It functions as a natural anticoagulant by greatly accelerating activation of protein C by thrombin. Using a direct gene screening strategy we identified a frameshift insertion mutation, insT 1689, in the thrombomodulin gene of a patient with myocardial infarction. The mutation predicts an elongated gene product because of substitution of the 12 C-terminal amino acids by 61 abnormal residues. Pedigree analysis showed that the mutation was also likely to have been present in a sibling who had had fatal myocardial infarction. Carriers of the mutant allele express significantly lower amounts of thrombomodulin on the surface of their monocytes detected by flow cytometry and have lower levels of soluble thrombomodulin in plasma. Wild type and the mutant thrombomodulin were expressed in COS-7 cells. Cellular distribution of the expressed proteins was evaluated by immunofluorescence microscopy, which showed reduced cell surface expression and intense juxtanuclear localization of the abnormal protein. This suggests impaired translocation through the endoplasmic reticulum/Golgi apparatus. Cells expressing abnormal thrombomodulin had reduced ability ( approximately 2.5-fold) to accelerate the thrombin mediated activation of protein C. This is the first demonstration of reduced expression arising from a natural thrombomodulin gene mutation. The results provide support for the suggestion that gene mutation of thrombomodulin may be important in the pathogenesis of some cases of occlusive thrombotic disease. (Blood. 2000;95:569-576)
authors.affiliation: Division of Investigative Science, National Heart and Lung Institute, Division of Biomedical Sciences, Imperial College School of Medicine, London.
authors.forename: G
authors.initials: G
authors.lastname: Kunz
authors.forename: H A
authors.initials: HA
authors.lastname: Ireland
authors.forename: P J
authors.initials: PJ
authors.lastname: Stubbs
authors.forename: M
authors.initials: M
authors.lastname: Kahan
authors.forename: G C
authors.initials: GC
authors.lastname: Coulton
authors.forename: D A
authors.initials: DA
authors.lastname: Lane
mesh_terms: D000328:Adult; D000368:Aged; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000911:Antibodies, Monoclonal; D001483:Base Sequence; D002784:Cholesterol / Q000097:blood; D004251:DNA Transposable Elements; D005260:Female; D016368:Frameshift Mutation; D005786:Gene Expression Regulation; D006801:Humans; D006973:Hypertension / Q000235:genetics; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D009203:Myocardial Infarction / Q000097:blood / Q000235:genetics*; D010375:Pedigree; D017354:Point Mutation; D018807:Polymorphism, Single-Stranded Conformational; D018180:Thrombomodulin / Q000097:blood / Q000737:chemistry / Q000235:genetics*; D014280:Triglycerides / Q000097:blood
chemical_list: D000911:Antibodies, Monoclonal; D004251:DNA Transposable Elements; D018180:Thrombomodulin; D014280:Triglycerides; D002784:Cholesterol