The crayfish plasma clotting protein: A vitellogenin-related protein responsible for clot formation in crustacean blood

Abstract

Coagulation in crayfish blood is based on the transglutaminase-mediated crosslinking of a specific plasma clotting protein. Here we report the cloning of the subunit of this clotting protein from a crayfish hepatopancreas cDNA library. The ORF encodes a protein of 1,721 amino acids, including a signal peptide of 15 amino acids. Sequence analysis reveals that the clotting protein is homologous to vitellogenins, which are proteins found in vitellogenic females of egg-laying animals. The clotting protein and vitellogenins are all lipoproteins and share a limited sequence similarity to certain other lipoproteins (e.g., mammalian apolipoprotein B and microsomal triglyceride transfer protein) and contain a stretch with similarity to the D domain of mammalian von Willebrand factor. The crayfish clotting protein is present in both sexes, unlike the female-specific vitellogenins. Electron microscopy was used to visualize individual clotting protein molecules and to study the transglutaminase-mediated clotting reaction. In the presence of an endogenous transglutaminase, the purified clotting protein molecules rapidly assemble into long, f lexible chains that occasionally branch.

Genome-wide analysis of the Drosophila immune response by using oligonucleotide microarrays

“…The coagulation process is well studied in other arthropods like Limulus and crayfish (20,23), but remains uncharacterized in Drosophila. This process involves serine protease processing of a clottable protein typically through a cascade.…”

“…This process involves serine protease processing of a clottable protein typically through a cascade. Curiously, the fly genome does not encode homologues of Limulus or crayfish clotting proteins (20,23), but encodes 10 fibrinogen-like proteins and several serine proteases that present significant homology with vertebrate coagulation factors. We found one fibrinogenlike encoding gene (CG5550) that is strongly induced after septic injury, representing an obvious candidate for the clottable protein in Drosophila.…”

A Cysteine-rich Protein from an Arthropod Stabilizes Clotting Mesh and Immobilizes Bacteria at Injury Sites

“…In mammals blood coagulation is based on the proteolytically induced polymerization of fibrinogen (2). Initially, fibrin monomers non-covalently interact with each other, and the resulting homopolymers are further stabilized by the plasma transglutaminase (TGase) 4 through the intermolecular cross-linking of ⑀-(␥ -glutamyl) lysine bonds (3). In crustaceans, although no proteolytic cascades are involved in the hemolymph coagulation, the coagulation also depends on the TGase-mediated cross-linking of specific plasma-clotting proteins (1,4).…”

Interaction of Pathogenic Vibrio Bacteria With the Blood Clot of the Pacific White Shrimp, Litopenaeus vannamei

“…The three taxa that have seen the best biochemical characterization of the clotting process are the vertebrates (Furie and Furie, 2000), the chelicerate arthropods (Muta and Iwanaga, 1996), and the crustaceans (Fuller and Doolittle, 1971a, b;Kopacek et al, 1993;Hall et al, 1999). In vertebrates and chelicerates, the initiation of the polymerization of the precursor structural protein into the fibrils of the clot involves the proteolytic activation of a soluble zymogen.…”

“…The protolyzed zymogens, fibrin in vertebrates (Weisel, 2005) and coagulin in chelicerates (Kawasaki et al, 2000), assemble into the clot fibrils. The crustacean clot is established by the transglutaminase-catalyzed covalent cross-linking of an abundant plasma lipoprotein, variously named clotting protein (CP) and very high density lipoprotein (VHDL) (Fuller and Doolittle, 1971a, b;Doolittle and Riley, 1990;Hall et al, 1999;Wang et al, 2001;Maningas et al, 2013).…”