Production, crystallization and preliminary crystallographic analysis of an exosite-containing fragment of human von Willebrand factor-cleaving proteinase ADAMTS13
Production, crystallization and preliminary crystallographic analysis of an exosite-containing fragment of human von Willebrand factor-cleaving proteinase ADAMTS13
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DOI：
10.1107/s1744309109023410
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发表时间：
2009-07-01
影响因子：
0.9
通讯作者：
Miyata, Toshiyuki
中科院分区：
文献类型：
--
作者：
Akiyama, Masashi;Takeda, Soichi;Miyata, Toshiyuki
ADAMTS 13是属于ADAMTS（具有血小板反应蛋白1型基序的去整合素和金属蛋白酶）家族的replysin型金属蛋白酶。它特异性地切割血浆血管性血友病因子（VWF）并调节血小板粘附和聚集。ADAMTS 13是一种多结构域酶。除了N-末端金属蛋白酶结构域之外，辅助结构域，包括去整合素样结构域、血小板反应蛋白-1 1型重复序列、富含Cys的结构域和间隔区结构域，是VWF识别和切割所需的。在本研究中，使用CHO Lec细胞表达ADAMTS 13辅助结构域的片段（ADAMTS 13-DTCS;残基287-685），并进行纯化和结晶。使用SPring-8光束线收集衍射数据集。两个ADAMTS 13-DTCS晶体与不同的晶胞参数产生的数据集，分别为2.6和2.8埃的分辨率。
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 angstrom resolution, respectively.