BIOMAT Database Logo BIOMAT Database Logo

Mutational analysis of antistasin, an inhibitor of blood coagulation factor Xa derived from the Mexican leech Haementeria officinalis. 1994

H J Theunissen, and R Dijkema, and J C Swinkels, and T L de Poorter, and P M Vink, and T G van Dinther
Department of Biotechnology and Biochemistry, N. V. Organon, Oss, The Netherlands.

Antistasin is a Factor Xa inhibitor that is present in the salivary glands of the Mexican leech Haementeria officinalis. The antistasin protein consists of 119 amino acids, of which residues 1-55 (domain I) are 56% similar to residues 56-110 (domain II). Of the nine C-terminal amino acids (residues 111-119; domain III), four are positively charged. The reactive site for Factor Xa is located in domain I. In this study we assessed the role of separate domains and of individual amino acids in the reactive site for the inhibition of Factor Xa. A series of mutants was constructed and expressed in Chinese hamster ovary (CHO) cells. In vitro chromogenic assays for Factor Xa show that domain I is sufficient for inhibition of Factor Xa. Domains II and III neither contain any intrinsic Factor Xa inhibitory activity, nor contribute to the activity of domain I. Furthermore, domain II does not become a Factor Xa inhibitor by partially adaptating its sequence towards that of the reactive site in domain I. Mutation of the cysteine at position 33 is not crucial for Factor Xa inhibition, suggesting a relatively rigid reactive site loop structure.

UI MeSH Term Description Entries
D007447 Invertebrate Hormones Hormones produced by invertebrates, usually insects, mollusks, annelids, and helminths. Hormones, Invertebrate
D007865 Leeches Annelids of the class Hirudinea. Some species, the bloodsuckers, may become temporarily parasitic upon animals, including man. Medicinal leeches (HIRUDO MEDICINALIS) have been used therapeutically for drawing blood since ancient times. Hirudinea,Hirudineas,Leeche
D008969 Molecular Sequence Data Descriptions of specific amino acid, carbohydrate, or nucleotide sequences which have appeared in the published literature and/or are deposited in and maintained by databanks such as GENBANK, European Molecular Biology Laboratory (EMBL), National Biomedical Research Foundation (NBRF), or other sequence repositories. Sequence Data, Molecular,Molecular Sequencing Data,Data, Molecular Sequence,Data, Molecular Sequencing,Sequencing Data, Molecular
D004252 DNA Mutational Analysis Biochemical identification of mutational changes in a nucleotide sequence. Mutational Analysis, DNA,Analysis, DNA Mutational,Analyses, DNA Mutational,DNA Mutational Analyses,Mutational Analyses, DNA
D006224 Cricetinae A subfamily in the family MURIDAE, comprising the hamsters. Four of the more common genera are Cricetus, CRICETULUS; MESOCRICETUS; and PHODOPUS. Cricetus,Hamsters,Hamster
D000595 Amino Acid Sequence The order of amino acids as they occur in a polypeptide chain. This is referred to as the primary structure of proteins. It is of fundamental importance in determining PROTEIN CONFORMATION. Protein Structure, Primary,Amino Acid Sequences,Sequence, Amino Acid,Sequences, Amino Acid,Primary Protein Structure,Primary Protein Structures,Protein Structures, Primary,Structure, Primary Protein,Structures, Primary Protein
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D000925 Anticoagulants Agents that prevent BLOOD CLOTTING. Anticoagulant Agent,Anticoagulant Drug,Anticoagulant,Anticoagulant Agents,Anticoagulant Drugs,Anticoagulation Agents,Indirect Thrombin Inhibitors,Agent, Anticoagulant,Agents, Anticoagulant,Agents, Anticoagulation,Drug, Anticoagulant,Drugs, Anticoagulant,Inhibitors, Indirect Thrombin,Thrombin Inhibitors, Indirect
D015342 DNA Probes Species- or subspecies-specific DNA (including COMPLEMENTARY DNA; conserved genes, whole chromosomes, or whole genomes) used in hybridization studies in order to identify microorganisms, to measure DNA-DNA homologies, to group subspecies, etc. The DNA probe hybridizes with a specific mRNA, if present. Conventional techniques used for testing for the hybridization product include dot blot assays, Southern blot assays, and DNA:RNA hybrid-specific antibody tests. Conventional labels for the DNA probe include the radioisotope labels 32P and 125I and the chemical label biotin. The use of DNA probes provides a specific, sensitive, rapid, and inexpensive replacement for cell culture techniques for diagnosing infections. Chromosomal Probes,DNA Hybridization Probe,DNA Probe,Gene Probes, DNA,Conserved Gene Probes,DNA Hybridization Probes,Whole Chromosomal Probes,Whole Genomic DNA Probes,Chromosomal Probes, Whole,DNA Gene Probes,Gene Probes, Conserved,Hybridization Probe, DNA,Hybridization Probes, DNA,Probe, DNA,Probe, DNA Hybridization,Probes, Chromosomal,Probes, Conserved Gene,Probes, DNA,Probes, DNA Gene,Probes, DNA Hybridization,Probes, Whole Chromosomal
D016466 CHO Cells CELL LINE derived from the ovary of the Chinese hamster, Cricetulus griseus (CRICETULUS). The species is a favorite for cytogenetic studies because of its small chromosome number. The cell line has provided model systems for the study of genetic alterations in cultured mammalian cells. CHO Cell,Cell, CHO,Cells, CHO

Related Publications

H J Theunissen, and R Dijkema, and J C Swinkels, and T L de Poorter, and P M Vink, and T G van Dinther
Purification and characterization of recombinant antistasin: a leech-derived inhibitor of coagulation factor Xa.
February 1991, Archives of biochemistry and biophysics,
H J Theunissen, and R Dijkema, and J C Swinkels, and T L de Poorter, and P M Vink, and T G van Dinther
Expression and characterization of the N-terminal half of antistasin, an anticoagulant protein derived from the leech Haementeria officinalis.
February 1991, Protein expression and purification,
H J Theunissen, and R Dijkema, and J C Swinkels, and T L de Poorter, and P M Vink, and T G van Dinther
A new factor Xa inhibitor (lefaxin) from the Haementeria depressa leech.
November 1999, Thrombosis and haemostasis,
H J Theunissen, and R Dijkema, and J C Swinkels, and T L de Poorter, and P M Vink, and T G van Dinther
Isolation and structural characterization of a potent inhibitor of coagulation factor Xa from the leech Haementeria ghilianii.
June 1989, Thrombosis and haemostasis,
H J Theunissen, and R Dijkema, and J C Swinkels, and T L de Poorter, and P M Vink, and T G van Dinther
Site-directed mutagenesis of the leech-derived factor Xa inhibitor antistasin. Probing of the reactive site.
November 1992, The Biochemical journal,
H J Theunissen, and R Dijkema, and J C Swinkels, and T L de Poorter, and P M Vink, and T G van Dinther
Antistasin, a leech-derived inhibitor of factor Xa. Kinetic analysis of enzyme inhibition and identification of the reactive site.
October 1989, The Journal of biological chemistry,
H J Theunissen, and R Dijkema, and J C Swinkels, and T L de Poorter, and P M Vink, and T G van Dinther
Functional characterisation of vizottin, the first factor Xa inhibitor purified from the leech Haementeria vizottoi.
September 2012, Thrombosis and haemostasis,
H J Theunissen, and R Dijkema, and J C Swinkels, and T L de Poorter, and P M Vink, and T G van Dinther
Tridegin, a new peptidic inhibitor of factor XIIIa, from the blood-sucking leech Haementeria ghilianii.
June 1997, The Biochemical journal,
H J Theunissen, and R Dijkema, and J C Swinkels, and T L de Poorter, and P M Vink, and T G van Dinther
Anticoagulant and antithrombotic action of the factor Xa inhibitor antistasin (ATS).
July 1993, Thrombosis research,
H J Theunissen, and R Dijkema, and J C Swinkels, and T L de Poorter, and P M Vink, and T G van Dinther
An inhibitor of collagen-stimulated platelet activation from the salivary glands of the Haementeria officinalis leech. I. Identification, isolation, and characterization.
April 1992, The Journal of biological chemistry,
Copied contents to your clipboard!