Coagulation factor XII protease domain crystal structure.
Coagulation factor XII protease domain crystal structure.
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DOI：
10.1111/jth.12849
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发表时间：
2015-04
期刊：
影响因子：
--
通讯作者：
Emsley J
中科院分区：
文献类型：
--
作者：
Pathak M;Wilmann P;Awford J;Li C;Hamad BK;Fischer PM;Dreveny I;Dekker LV;Emsley J
凝血因子XII是一种丝氨酸蛋白酶，其对于激肽生成和血液凝固很重要，切割底物血浆激肽释放酶和FXI。通过测定FXII蛋白酶结构域的晶体结构，研究FXII酶原激活和底物识别。通过测量显色肽和血浆激肽释放酶蛋白底物的切割来表征一系列重组FXII蛋白酶构建体。这表明，与β-FXIIa相比，跨越轻链的FXII蛋白酶构建体具有意想不到的较弱的蛋白水解活性，β-FXIIa具有额外的9个重链氨基酸残基。与这些数据一致，轻链蛋白酶的晶体结构揭示了活性位点残基Gly 193和Ser 195的酶原构象，其中不存在氧阴离子空穴。Asp 194侧链盐桥Arg 73构成了一个非典型的构象的70环。在一种晶体形式中，S1口袋环是部分柔性的，这是典型的酶原。在去糖基化轻链的第二种晶型中，S1口袋环是有序的，结构的180环中的短α螺旋导致具有Asp 189掩埋构象的扩大和扭曲的S1口袋，这对于P1 Arg底物识别至关重要。FXII结构定义了活性位点裂缝周围的负电荷补丁，这可能是与抑制剂和底物相互作用的关键。这些数据为理解FXII底物识别和酶原激活提供了第一个结构基础。
Coagulation factor XII is a serine protease that is important for kinin generation and blood coagulation, cleaving the substrates plasma kallikrein and FXI. To investigate FXII zymogen activation and substrate recognition by determining the crystal structure of the FXII protease domain. A series of recombinant FXII protease constructs were characterized by measurement of cleavage of chromogenic peptide and plasma kallikrein protein substrates. This revealed that the FXII protease construct spanning the light chain has unexpectedly weak proteolytic activity compared to β-FXIIa, which has an additional nine amino acid remnant of the heavy chain present. Consistent with these data, the crystal structure of the light chain protease reveals a zymogen conformation for active site residues Gly193 and Ser195, where the oxyanion hole is absent. The Asp194 side chain salt bridge to Arg73 constitutes an atypical conformation of the 70-loop. In one crystal form, the S1 pocket loops are partially flexible, which is typical of a zymogen. In a second crystal form of the deglycosylated light chain, the S1 pocket loops are ordered, and a short α-helix in the 180-loop of the structure results in an enlarged and distorted S1 pocket with a buried conformation of Asp189, which is critical for P1 Arg substrate recognition. The FXII structures define patches of negative charge surrounding the active site cleft that may be critical for interactions with inhibitors and substrates. These data provide the first structural basis for understanding FXII substrate recognition and zymogen activation.
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DOI：
10.1084/jem.20052458
发表时间：
2006-03-20
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Kleinschnitz C;Stoll G;Bendszus M;Schuh K;Pauer HU;Burfeind P;Renné C;Gailani D;Nieswandt B;Renné T
通讯作者：
Renné T
DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
Emsley, P;Cowtan, K
通讯作者：
Cowtan, K
影响因子：
17.1
作者：
Larsson, Magnus;Rayzman, Veronika;Renne, Thomas
通讯作者：
Renne, Thomas
影响因子：
5.6
作者：
Shia, S;Stamos, J;Eigenbrot, C
通讯作者：
Eigenbrot, C
影响因子：
7.5
作者：
HOJIMA, Y;TANKERSLEY, DL;PISANO, JJ
通讯作者：
PISANO, JJ