Open AccessCloning and expression of a cDNA coding for the anticoagulant hirudin from the bloodsucking leech, Hirudo medicinalis.
Author(s) -
Richard P. Harvey,
Eric Degryse,
Lenita M. Stefani,
Fabienne Schamber,
JeanPierre Cazenave,
Michael Courtney,
Paul Tolstoshev,
JeanPierre Lecocq
Publication year - 1986
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.83.4.1084
Subject(s) - hirudo medicinalis , hirudin , leech , complementary dna , biology , biochemistry , thrombin , molecular cloning , microbiology and biotechnology , peptide sequence , glycoprotein , recombinant dna , gene , platelet , world wide web , computer science , immunology
Cloned cDNAs have been isolated that encode a variant of hirudin, a potent thrombin inhibitor that is secreted by the salivary glands of the medicinal leech, Hirudo medicinalis. This variant probably corresponds to a form that has been purified from leech heads but differs in amino acid sequence from the hirudin purified from whole leeches. There are at least three hirudin transcripts detectable in leech RNAs that are different in size, site of synthesis, inducibility by starvation, and relationship to hirudin activity. The new hirudin variant predicted by the cDNA and the heterodisperse transcription products suggest a hirudin protein family. The hirudin cDNA was expressed in Escherichia coli under the control of the bacteriophage lambda PL promoter. The recombinant product is biologically active, inhibiting the cleavage by thrombin of fibrinogen and a synthetic tripeptide substrate.