title: The spider hemolymph clot proteome reveals high concentrations of hemocyanin and von Willebrand factor-like proteins.
abstract: Arthropods include chelicerates, crustaceans, and insects that all have open circulation systems and thus require different properties of their coagulation system than vertebrates. Although the clotting reaction in the chelicerate horseshoe crab (Family: Limulidae) has been described in details, the overall protein composition of the resulting clot has not been analyzed for any of the chelicerates. The largest class among the chelicerates is the arachnids, which includes spiders, ticks, mites, and scorpions. Here, we use a mass spectrometry-based approach to characterize the spider hemolymph clot proteome from the Brazilian whiteknee tarantula, Acanthoscurria geniculata. We focused on the insoluble part of the clot and demonstrated high concentrations of proteins homologous to the hemostasis-related and multimerization-prone von Willebrand factor. These proteins, which include hemolectins and vitellogenin homologous, were previously identified as essential components of the hemolymph clot in crustaceans and insects. Their presence in the spider hemolymph clot suggests that the origin of these proteins' function in coagulation predates the split between chelicerates and mandibulata. The clot proteome reveals that the major proteinaceous component is the oxygen-transporting and phenoloxidase-displaying abundant hemolymph protein hemocyanin, suggesting that this protein also plays a role in clot biology. Furthermore, quantification of the peptidome after coagulation revealed the simultaneous activation of both the innate immune system and the coagulation system. In general, many of the identified clot-proteins are related to the innate immune system, and our results support the previously suggested crosstalk between immunity and coagulation in arthropods.
authors.affiliation: Interdisciplinary Nanoscience Center and Department of Molecular Biology and Genetics, Aarhus University, 8000 Aarhus C, Denmark.
authors.forename: Kristian W
authors.initials: KW
authors.lastname: Sanggaard
authors.affiliation: Interdisciplinary Nanoscience Center and Department of Molecular Biology and Genetics, Aarhus University, 8000 Aarhus C, Denmark.
authors.forename: Thomas F
authors.initials: TF
authors.lastname: Dyrlund
authors.affiliation: Department of Bioscience, Aarhus University, 8000 Aarhus C, Denmark.
authors.forename: Jesper S
authors.initials: JS
authors.lastname: Bechsgaard
authors.affiliation: Interdisciplinary Nanoscience Center and Department of Molecular Biology and Genetics, Aarhus University, 8000 Aarhus C, Denmark.
authors.forename: Carsten
authors.initials: C
authors.lastname: Scavenius
authors.affiliation: Department of Bioscience, Aarhus University, 8000 Aarhus C, Denmark.
authors.forename: Tobias
authors.initials: T
authors.lastname: Wang
authors.affiliation: Department of Bioscience, Aarhus University, 8000 Aarhus C, Denmark.
authors.forename: Trine
authors.initials: T
authors.lastname: Bilde
authors.affiliation: Interdisciplinary Nanoscience Center and Department of Molecular Biology and Genetics, Aarhus University, 8000 Aarhus C, Denmark. Electronic address: jje@mbg.au.dk.
authors.forename: Jan J
authors.initials: JJ
authors.lastname: Enghild
mesh_terms: D000818:Animals; D001777:Blood Coagulation / Q000235:genetics; D001938:Brazil; D006433:Hemocyanins / Q000096:biosynthesis* / Q000235:genetics; D006458:Hemolymph / Q000378:metabolism*; D020543:Proteome / Q000235:genetics*; D013112:Spiders / Q000235:genetics / Q000378:metabolism; D014841:von Willebrand Factor / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D020543:Proteome; D014841:von Willebrand Factor; D006433:Hemocyanins
keywords: Arthropods; Coagulation; Evolution; Hemocyanin; Innate immunity; Proteomics