PMID: 3768302Aug 12, 1986Paper

Kinetics of the inhibition of thrombin by hirudin

Biochemistry
S R Stone, J Hofsteenge

Abstract

The dissociation constant for hirudin was determined by varying the concentration of hirudin in the presence of a fixed concentration of thrombin and tripeptidyl p-nitroanilide substrate. The estimate of the dissociation constant determined in this manner displayed a dependence on the concentration of substrate which suggested the existence of two binding sites at which the substrate was able to compete with hirudin. A high-affinity site could be correlated with the binding of the substrate at the active site, and the other site had an affinity for the substrate that was 2 orders of magnitude lower. Extrapolation to zero substrate concentration yielded a value of 20 fM for the dissociation constant of hirudin at an ionic strength of 0.125. The dissociation constant for hirudin was markedly dependent on the ionic strength of the assay; it increased 20-fold when the ionic strength was increased from 0.1 to 0.4. This increase in dissociation constant was accompanied by a decrease in the rate with which hirudin associated with thrombin. This rate could be measured with a conventional recording spectrophotometer at higher ionic strength and was found to be independent of the binding of substrate at the active site.

References

Jun 12, 1979·Biochemistry·J W WilliamsR G Duggleby
Jan 1, 1979·Methods in Enzymology·J W Williams, J F Morrison
Jan 1, 1976·Methods in Enzymology·D BagdyS Magnusson
Jun 1, 1975·Archives of Biochemistry and Biophysics·W J Landis, D F Waugh
Jul 29, 1965·Biochimica Et Biophysica Acta·M J Selwyn
Jan 1, 1984·Molecular and Cellular Biochemistry·L J Berliner
Feb 15, 1983·Biochimica Et Biophysica Acta·R Lottenberg, C M Jackson
Jan 1, 1981·Annals of the New York Academy of Sciences·J W Fenton
May 14, 1982·Biochemical and Biophysical Research Communications·E Van Obberghen-SchillingJ Pouysségur
Jan 1, 1982·Methods in Enzymology·K J Ellis, J F Morrison
Jan 1, 1981·Analytical Biochemistry·R G Duggleby

❮ Previous
Next ❯

Citations

Jun 25, 1999·Biopolymers·A LombardiV Pavone
Jan 1, 1994·Journal of Clinical Pharmacology·L A Harker
May 1, 1993·Medicinal Research Reviews·R A Wiley, D H Rich
Jun 1, 1992·Protein Science : a Publication of the Protein Society·A KarshikovS R Stone
Feb 1, 1993·Protein Science : a Publication of the Protein Society·G L Rosenquist, H B Nicholas
Oct 1, 1993·Protein Science : a Publication of the Protein Society·J P PriestleM G Grütter
Feb 1, 1994·Protein Science : a Publication of the Protein Society·U Neumann, J Hofsteenge
Jun 1, 1996·Protein Science : a Publication of the Protein Society·J FéthièreM Cygler
Apr 1, 1998·Protein Science : a Publication of the Protein Society·G De SimoneV Pavone
Jan 1, 1995·Journal of Thrombosis and Thrombolysis·C P Cannon
Aug 1, 1991·Annals of Hematology·J Bichler, H Fritz
Jun 2, 1989·Journal of Immunological Methods·S J MaoJ L Krstenansky
Sep 20, 1991·Journal of Molecular Biology·T J RydelR Huber
Dec 1, 1988·Trends in Pharmacological Sciences·R B Wallis
Oct 1, 1993·Trends in Pharmacological Sciences·C TapparelliN S Cook
May 18, 1995·Biochimica Et Biophysica Acta·G SanyalC R Middaugh
Aug 30, 1996·Biophysical Chemistry·K A Sharp
Jun 1, 1995·Journal of the American College of Cardiology·C P Cannon, E Braunwald
Aug 1, 1995·Bioorganic & Medicinal Chemistry·J Das, S D Kimball
Sep 1, 1992·Trends in Cardiovascular Medicine·N U Bang, M D Clayman
Jun 2, 2005·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Mercedes López, Goetz Nowak
Nov 11, 2003·Trends in Pharmacological Sciences·James A Huntington, Trevor P Baglin
Mar 4, 2005·The American Journal of Pathology·Shu Jun SongEleanor J Mackie
Jun 8, 2002·Journal of Molecular Biology·Bernhard SchlottOliver Ohlenschläger
Oct 24, 2002·Bioorganic Chemistry·Brian E CathersRobert M Rydzewski
Apr 18, 2002·Thrombosis Research·Sedat KirazMeral Calgüneri
Jun 24, 2003·Thrombosis Research·Jack Hirsh
Nov 15, 1996·Thrombosis Research·J BichlerW G Owen
Jan 15, 1997·Thrombosis Research·A C Teger-NilssonU Eriksson
Apr 2, 1998·General Pharmacology·P Ramaprasad, C Kasinathan
Apr 2, 1998·General Pharmacology·M CappielloU Mura

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.