title: Purification and partial characterization of the plasma clotting protein from the pink shrimp Farfantepenaeus paulensis.
abstract: A clotting protein (CP) was purified from the plasma of the pink shrimp Farfantepenaeus paulensis by sequential anion-exchange chromatography. The shrimp CP was able to form stable clots in vitro in the presence of hemocyte lysate and Ca2+, suggesting that the clotting reaction is catalyzed by a Ca2+-dependent transglutaminase present in shrimp hemocytes. Dansylcadaverine was incorporated into the shrimp CP in the presence of endogenous transglutaminase (hemocyte lysate), confirming that the shrimp purified CP is the substrate for the transglutaminase enzyme. The molecular mass of the CP was determined by gel filtration to be 341 kDa and 170 kDa by SDS-PAGE under reducing conditions. These results suggest that the shrimp CP consists of two identical subunits, covalently linked by disulphide bonds. The amino acid sequence at the N-terminus was 100% identical to that of the penaeids Litopenaeus vannamei and Penaeus monodon and 66% to 80% identical to the CPs of other decapods. This is the first report of a CP characterization in an Atlantic penaeid species. Further studies, including a molecular cloning approach would enable to detect which tissues express the gene of the clotting protein. It would be also useful to understand the mechanism by which the coagulation time is delayed in shrimps under stress conditions.
authors.affiliation: Laboratório de Imunologia Aplicada à Aqüicultura, Departamento de Biologia Celular, Embriologia e Genética (BEG), Centro de Ciências Biológicas (CCB), Universidade Federal de Santa Catarina (UFSC), CP. 476, CEP 88040-900, Florianópolis, SC, Brazil.
authors.forename: Luciane M
authors.initials: LM
authors.lastname: Perazzolo
authors.forename: Daniel M
authors.initials: DM
authors.lastname: Lorenzini
authors.forename: Sirlei
authors.initials: S
authors.lastname: Daffre
authors.forename: Margherita A
authors.initials: MA
authors.lastname: Barracco
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001777:Blood Coagulation / Q000502:physiology*; D001798:Blood Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D002103:Cadaverine / Q000031:analogs & derivatives / Q000378:metabolism; D005260:Female; D005456:Fluorescent Dyes / Q000378:metabolism; D008074:Lipoproteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D008297:Male; D008969:Molecular Sequence Data; D008970:Molecular Weight; D033561:Penaeidae / Q000378:metabolism*; D021122:Protein Subunits / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D016415:Sequence Alignment; D011503:Transglutaminases / Q000037:antagonists & inhibitors / Q000378:metabolism
chemical_list: D001798:Blood Proteins; D005456:Fluorescent Dyes; D008074:Lipoproteins; D021122:Protein Subunits; D011503:Transglutaminases; C008542:monodansylcadaverine; D002103:Cadaverine