title: [Research on the C-terminal domain of ADAMTS13 regulates its cleaving activity].
abstract: OBJECTIVE
  To study the influence of C-terminal domain of ADAMTS13 on its cleaving activity.
  
  
  METHODS
  The full-length wild-type (WT) and C-terminal domain truncated type (TT, TSP8 + CUB domains were deleted) of human ADAMTS13 recombinant protein were transfected into and permanent expressed on Hela cells. Western blot and R-CBA were used to directly detect the activities of the two recombinant proteins under the static and stressed condition respectively. ELISA was used to compare the binding abilities of the two proteins by coating with vWF.
  
  
  RESULTS
  The recombinant proteins were identified by Western blot with anti-his-tag or anti-ADAMTS13 antibodies. With pretreatment of 1.5 M urea, the enzyme activity of TT was significantly higher than that of WT, and so did in binding ability with vWF While, only WT could cleave vWF under high stress.
  
  
  CONCLUSIONS
  The distal carboxyl-terminal TSP8 together with CUB domains of ADAMTS13 may affect the enzyme activity by regulating the binding of ADAMTS13 to vWF in different conditions, and they are very important for the enzyme activity under high stress force condition.
authors.affiliation: Jiangsu Institute of Hematology, The First Affiliated Hospital of Soochow University, Key Laboratory of Thrombosis and Hemostasis of Ministry of Health, Suzhou 215006, China.
authors.forename: An-You
authors.initials: AY
authors.lastname: Wang
authors.forename: Fang
authors.initials: F
authors.lastname: Liu
authors.forename: Zhen-Ni
authors.initials: ZN
authors.lastname: Ma
authors.forename: Ning-Zheng
authors.initials: NZ
authors.lastname: Dong
authors.forename: Jing-Yu
authors.initials: JY
authors.lastname: Zhang
authors.forename: Chang-Geng
authors.initials: CG
authors.lastname: Ruan
mesh_terms: D032061:Galium; D006801:Humans; D011994:Recombinant Proteins / Q000378:metabolism; D014162:Transfection; D014841:von Willebrand Factor / Q000235:genetics
chemical_list: D011994:Recombinant Proteins; D014841:von Willebrand Factor