Genetic analysis and functional characterization of Prothrombins Corpus Christi (Arg382-cys), Dhahran (Arg271-His), and hypoprothrombinemia

• Published in: Blood Vol. 88; no. 7; pp. 2611 - 2618
• Main Authors: O'MARCIAGH, A. S, NICHOLS, W. L, HASSINGER, N. L, MULLINS, J. D, MALLOUH, A. A, GILCHRIST, G. S, OWEN, W. G
• Format: Journal Article
• Language: English
• Published: Washington, DC The Americain Society of Hematology 01.10.1996
• Subjects: Adult; Amino Acid Sequence; Base Sequence; Biological and medical sciences; Child; Child, Preschool; Codon - genetics; DNA Mutational Analysis; Erythema - genetics; Factor Xa - metabolism; Female; Genotype; Hematologic and hematopoietic diseases; Hemorrhagic Disorders - genetics; Humans; Hypoprothrombinemias - genetics; Male; Medical sciences; Middle Aged; Partial Thromboplastin Time; Pedigree; Platelet Aggregation; Platelet diseases and coagulopathies; Polymerase Chain Reaction; Prothrombin - chemistry; Prothrombin - genetics; Prothrombin - physiology; Prothrombin Time; Structure-Activity Relationship; Thrombin - physiology; Human; Variant; Blood coagulation; Nucleotide sequence; Family study; Fibrinogen; Prothrombin; Mutation; Genome; Mechanism of action; Biological activity
• ISSN: 0006-4971; 1528-0020
• DOI: 10.1182/blood.v88.7.2611.bloodjournal8872611

The structural abnormalities and functional characteristics of dysfunctional prothrombin variants in two new kindreds have been determined. Prothrombin Corpus Christi (family 1) was purified and found to have markedly reduced fibrinogen clotting activity, yet normal amidolytic and near-normal platelet aggregating activity. A transition (C to T) at nucleotide position 8885, present in the heterozygous form in affected family members, resulted in the substitution of Cys for Arg 382. This substitution results in the loss of a positive charge within the fibrinogen-binding exosite of thrombin, thus accounting for the observed functional defect. A heterozygous C to T transition was also present at position 19994 in other family members with a hypoprothrombinemic phenotype. This mutation results in the replacement of Gln 541 (CAA) by a premature stop codon (TAA). Prothrombin Dhahran (family 2) was found to have markedly reduced fibrinogen clotting activity, but normal amidolytic activity. Affected family members were found to have a G to A transition at nucleotide position 7312 resulting in the substitution of His for Arg 271. This substitution results in the abolition of a factor Xa cleavage site, yielding meizothrombin rather than thrombin, on activation of prothrombin Dhahran by factor Xa. All but one of the above mutations occur at CpG dinucleotides, thus further supporting the observation of a high incidence of CpG transitions in hereditary dysprothrombinemia. The significant bleeding tendencies of individuals homozygous for prothrombin Dhahran (prothrombin clotting activity 5% to 7%) contrast sharply with the absence of significant chronic bleeding in the proband expressing prothrombin Corpus Christi (prothrombin clotting activity 2%). Our findings underscore the capacity of thrombin to contribute to clinical hemostasis by mechanisms other than its fibrinogen clotting activity.