title: Isolation, sequence analysis, and cloning of haemadin. An anticoagulant peptide from the Indian leech.
abstract: A slow, tight-binding inhibitor of thrombin with an apparent molecular mass of about 5 kDa has been isolated from Haemadipsa sylvestris, an Indian leech of the family of Haemadipsidae. The inhibitory activity, called haemadin, is thrombin specific since it does not inhibit other proteases like trypsin, chymotrypsin, factor Xa, or plasmin. NH2-terminal amino acid sequence analysis (residues 1-45) does not reveal any homology to known serine protease inhibitors, including the thrombin-specific inhibitor hirudin. The haemadin cDNA cloned by polymerase chain reaction techniques codes for a polypeptide of 57 amino acid residues preceded by 20 residues of a signal peptide sequence. A synthetic gene coding for the mature haemadin was expressed in Escherichia coli. Recombinant haemadin displays a similar inhibition constant and specific activity as its natural counterpart. Although there is no obvious sequence identity between haemadin and hirudin, both proteins seem to share common mechanisms for thrombin inhibition.
authors: Department of Biotechnology, BASF Aktiengesellschaft, Ludwigshafen, Germany.
authors: K H
authors: KH
authors: Strube
authors: B
authors: B
authors: Kröger
authors: S
authors: S
authors: Bialojan
authors: M
authors: M
authors: Otte
authors: J
authors: J
authors: Dodt
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000925:Anticoagulants / Q000737:chemistry / Q000302:isolation & purification*; D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D004247:DNA; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli; D006801:Humans; D007447:Invertebrate Hormones / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D007700:Kinetics; D007865:Leeches / Q000737:chemistry*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D013917:Thrombin / Q000037:antagonists & inhibitors*
chemical_list: D000925:Anticoagulants; D007447:Invertebrate Hormones; C080680:haemadin; D004247:DNA; D013917:Thrombin