title: Sea urchin coelomocyte arylsulfatase: a modulator of the echinoderm clotting pathway.
abstract: Sea urchin petalloid coelomocytes effectuate the clotting pathway by undergoing a rapid and dynamic cellular transformation that leads to cellular adhesion and wounds closure. We have identified high levels of activity of arylsulfatase (Ars) associated with coelomocytes of the sea urchin Lytechinus variegatus (Lamarck, 1816). Ars activity was extracted from clotted coelomocytes with EDTA and showed high levels of activity up to a 1:100 dilution. Clot formation from isolated coelomic fluid was significantly inhibited by the ARS inhibitor, p-nitrophenyl phosphate. Ars activity was collected by 80% ethanol precipitation, a diagnostic test previously used in Ars isolation. Cellular extraction studies in the presence and absence of the non-ionic detergent Triton X-100 indicated that some Ars activity was present intracellularly, possibly in intracellular membrane-bound compartments, however the majority of Ars activity was extracted from the extracellular coelomocyte membrane. Polyclonal anti-sea urchin embryo Ars antibodies recognized a single protein band with an approximate molecular weight of 75 kDa on western blots. Immunofluorescence using the anti-sea urchin Ars antibody revealed an intracellular and extracellular staining of Ars in both petalloid and filopodial coelomocytes. Taken together, these data indicate that coelomocyte Ars might be involved in cell-to-cell crosslinking of surface sulfated polysaccharides vital for clot formation.
authors.affiliation: Department of Biology and Biochemistry, Northwestern College, St. Paul, Minnesota, USA. ldwinslow@nwc.edu
authors.forename: Lisanne
authors.initials: L
authors.lastname: D'Andrea-Winslow
authors.forename: David W
authors.initials: DW
authors.lastname: Radke
authors.forename: Tim
authors.initials: T
authors.lastname: Utecht
authors.forename: Takuya
authors.initials: T
authors.lastname: Kaneko
authors.forename: Koji
authors.initials: K
authors.lastname: Akasaka
mesh_terms: D000818:Animals; D001192:Arylsulfatases / Q000235:genetics / Q000378:metabolism*; D002448:Cell Adhesion; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology*; D047331:Lytechinus / Q000166:cytology*; D014945:Wound Healing
chemical_list: D001192:Arylsulfatases