Theromin, a novel leech thrombin inhibitor.
We purified the most potent thrombin inhibitor described to date from the rhynchobdellid leech Theromyzon tessulatum. Designated theromin, it was purified to apparent homogeneity by gel permeation and anion exchange chromatography followed by two reverse-phase steps of high performance liquid chromatography. The primary sequence of theromin (a homodimer of 67 amino acid residues including 16 cysteine residues) was determined by a combination of reduction and s-beta-pyridylethylation, Edman degradation, trypsin enzymatic digestion, and matrix-assisted laser desorption mass spectrometry measurement. Theromin exhibits no sequence homology with any other thrombin inhibitors. Furthermore, theromin significantly diminishes, in a dose-dependent manner, the level of human granulocyte and monocyte activation induced by lipopolysaccharides. In summary, this potent thrombin inhibitor promises to have high biomedical significance.
Salzet
M
M
Laboratoire d'Endocrinologie des Annélides, Unité Propre de la Recherche Supérieure Associée au CNRS 8017 CNRS, SN3, Université des Sciences et Technologie de Lille, F-59655 Villeneuve d'Ascq Cedex, France. michel.salzet@univ-lille1.fr
Chopin
V
V
Baert
J
J
Matias
I
I
Malecha
J
J
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000737:chemistry; D000818:Animals; D007611:Aprotinin / Q000494:pharmacology; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D006098:Granulocytes / Q000187:drug effects / Q000378:metabolism; D007700:Kinetics; D007865:Leeches; D008070:Lipopolysaccharides / Q000378:metabolism; D008213:Lymphocyte Activation / Q000187:drug effects; D008969:Molecular Sequence Data; D009000:Monocytes / Q000187:drug effects / Q000378:metabolism; D010455:Peptides / Q000737:chemistry; D011506:Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology / Q000502:physiology; D017386:Sequence Homology, Amino Acid; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013917:Thrombin / Q000037:antagonists & inhibitors*; D013997:Time Factors
D000596:Amino Acids; D008070:Lipopolysaccharides; D010455:Peptides; D011506:Proteins; C116698:tessulin; C415362:theromin; D007611:Aprotinin; D013917:Thrombin