+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Interaction of hirudin with the dysthrombins Quick I and II



Interaction of hirudin with the dysthrombins Quick I and II



Biochemistry 30(26): 6392-6397



The interaction of hirudin with the dysfunctional enzymes thrombin Quick I and II has been investigated. Natural and recombinant hirudin caused nonlinear competitive inhibition of thrombin Quick I. The results were consistent with thrombin Quick I existing in two forms that have different affinities for hirudin. The affinities of these forms for natural hirudin were respectively 10(4)- and 10(6)-fold lower than that of alpha-thrombin. In contrast, truncated hirudin molecules lacking the C-terminal tail of the molecule caused linear inhibition of thrombin Quick I. These results indicate that different modes of interaction of the two forms of thrombin Quick I with the C-terminal tail of hirudin were the cause of the nonlinear inhibition. Comparison of the dissociation constants of thrombin Quick I with the truncated and full-length forms of hirudin suggested that the interactions that normally occur between the C-terminal tail of hirudin and thrombin were completely disrupted with the low-affinity form of thrombin Quick I. Thrombin Quick II displayed an affinity for natural hirudin that was 10(3)-fold lower than that observed with alpha-thrombin. In contrast, it bound a mutant hirudin with altered N-terminal amino acids only 16-fold less tightly. These results are discussed in terms of structural alterations in the active-site cleft in thrombin Quick II.

Please choose payment method:






(PDF emailed within 0-6 h: $19.90)

Accession: 040466472

Download citation: RISBibTeXText

PMID: 2054344

DOI: 10.1021/bi00240a007


Related references

Inhibition of dysthrombins Quick I and II by heparin cofactor II and antithrombin. Journal of Biological Chemistry 268(5): 3321-3327, 1993

Preparation of monoclonal antibodies to hirudin and hirudin peptides. A method for studying the hirudin--thrombin interaction. European Journal of Biochemistry 188(2): 463-470, 1990

Effects of site specific amino acid exchanges in hirudin on the thrombin-hirudin interaction. Folia Haematologica 115(1-2): 24-29, 1988

Interaction of hirudin with thrombin identification of a minimal binding domain of hirudin that inhibits clotting activity. Biochemistry 27(21): 8170-8173, 1988

Use of fragments of hirudin to investigate thrombin-hirudin interaction. European Journal of Biochemistry 188(1): 61-66, 1990

Functional DNA block and plasmid coding for hirudin, transformed yeast, method for hirudin, hirudin obtained, and its pharmaceutical use. Official Gazette of the United States Patent and Trademark Office Patents 1222(2), 1999

Functional DNA block and plasmid coding for hirudin, transformed yeast, method for preparing hirudin, hirudin obtained and its pharmaceutical use. Official Gazette of the United States Patent and Trademark Office Patents 1194(1): 410-411, 1997

Anti-r-hirudin antibodies reveal clinical relevance through direct functional inactivation of r-hirudin or prolongation of r-hirudin's plasma halflife. Thrombosis and Haemostasis 85(5): 936-938, 2001

Mechanisms of the inhibition of alpha thrombin by hirudin derived fragments hirudin 1 47 and hirudin 45 65. European Journal of Biochemistry 195(1): 251-256, 1991

Capillary electrophoresis of r-hirudin and a polyethylene glycol derivative of r-hirudin (PEG-hirudin). Seminars in Thrombosis and Hemostasis 23(1): 39-43, 1997

Use of protein chemistry and molecular biology to determine interaction areas between proteases and their inhibitors the thrombin hirudin interaction as an example. Festoff, B W (Ed ) Nato Asi (Advanced Science Institutes) Series Series A Life Sciences, Vol 191 Serine Proteases And Their Serpin Inhibitors in The Nervous System: Regulation in Development And in Degenerative And Malignant Disease; Workshop, Maratea, Italy, July 2-8, 1989 Xxv+359p Plenum Publishing Corp : New York, New York, Usa; London, England, Uk Illus 115-126, 1990

A quick assay for monitoring recombinant hirudin during cardiopulmonary bypass in patients with heparin-induced thrombocytopenia type II : adaptation of the ecarin clotting time to the act II device. Journal of Thoracic and Cardiovascular Surgery 119(6): 1278-1283, 2000

Determination of r DNA hirudin and A-human thrombin- hirudin complex in plasma samples: enzyme linked immunosorbent assays for hirudin and complex vs. chromogenic thrombin substrate assay. Thrombosis Research 66(1): 33-42, 1992

PH dependence of the interaction of hirudin with thrombin. Biochemistry 31(4): 1168-1172, 1992