title: Tensilin-like stiffening protein from Holothuria leucospilota does not induce the stiffest state of catch connective tissue.
abstract: The dermis of sea cucumbers is a catch connective tissue or mutable connective tissue that exhibits large changes in mechanical properties. A stiffening protein, tensilin, has been isolated from the sea cucumber Cucumaria frondosa. We purified a similar protein, H-tensilin, from Holothuria leucospilota, which belongs to a different family to C. frondosa. H-tensilin appeared as a single band with an apparent molecular mass of 34 kDa on SDS-PAGE. No sugar chain was detected. Tryptic fragments of the protein had homology to known tensilin. H-tensilin aggregated isolated collagen fibrils in vitro in a buffer containing 0.5 mol l(-1) NaCl with or without 10 mmol l(-1) Ca(2+). The activity of H-tensilin was quantitatively studied by dynamic mechanical tests on the isolated dermis. H-tensilin increased stiffness of the dermis in the soft state, induced by Ca(2+)-free artificial seawater, to a level comparable to that of the standard state, which was the state found in the dermis rested in artificial seawater with normal ionic condition. H-tensilin decreased the energy dissipation ratio of the soft dermis to a level comparable to that of the standard state. When H-tensilin was applied on the dermis in the standard state, it did not alter stiffness nor dissipation ratio. The subsequent application of artificial seawater in which the potassium concentration was raised to 100 mmol l(-1) increased stiffness by one order of magnitude. These findings suggest that H-tensilin is involved in the changes from the soft state to the standard state and that some stiffening factors other than tensilin are necessary for the changes from the standard to the stiff state.
authors.affiliation: W3-42, Department of Biological Sciences, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, O-okayama 2-12-1, Tokyo 152-8551, Japan. mtamori@bio.titech.ac.jp
authors.forename: Masaki
authors.initials: M
authors.lastname: Tamori
authors.forename: Akira
authors.initials: A
authors.lastname: Yamada
authors.forename: Naoto
authors.initials: N
authors.lastname: Nishida
authors.forename: Yumiko
authors.initials: Y
authors.lastname: Motobayashi
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Oiwa
authors.forename: Tatsuo
authors.initials: T
authors.lastname: Motokawa
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002118:Calcium / Q000737:chemistry; D003238:Connective Tissue / Q000502:physiology*; D020405:Dermis / Q000502:physiology; D047332:Holothuria / Q000502:physiology*; D011188:Potassium; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D012623:Seawater
chemical_list: D011506:Proteins; D011188:Potassium; D002118:Calcium