Open AccessProduction, crystallization and preliminary crystallographic analysis of an exosite‐containing fragment of human von Willebrand factor‐cleaving proteinase ADAMTS13
Author(s) -
Akiyama Masashi,
Takeda Soichi,
Kokame Koichi,
Takagi Junichi,
Miyata Toshiyuki
Publication year - 2009
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309109023410
Subject(s) - adamts13 , thrombospondin , disintegrin , von willebrand factor , metalloproteinase , adamts , chemistry , cleavage (geology) , proteolysis , microbiology and biotechnology , biochemistry , biology , platelet , matrix metalloproteinase , enzyme , immunology , paleontology , fracture (geology)
ADAMTS13 is a reprolysin‐type metalloproteinase belonging to the ADAMTS (a disintegrin and metalloproteinase with thrombospondin type 1 motif) family. It specifically cleaves plasma von Willebrand factor (VWF) and regulates platelet adhesion and aggregation. ADAMTS13 is a multi‐domain enzyme. In addition to the N‐terminal metalloproteinase domain, the ancillary domains, including a disintegrin‐like domain, a thrombospondin‐1 type 1 repeat, a Cys‐rich domain and a spacer domain, are required for VWF recognition and cleavage. In the present study, a fragment of the ADAMTS13 ancillary domains (ADAMTS13‐DTCS; residues 287–685) was expressed using CHO Lec cells, purified and crystallized. Diffraction data sets were collected using the SPring‐8 beamline. Two ADAMTS13‐DTCS crystals with distinct unit‐cell parameters generated data sets to 2.6 and 2.8 Å resolution, respectively.