The serine protease domain of MASP-3: enzymatic properties and crystal structure in complex with ecotin.
The serine protease domain of MASP-3: enzymatic properties and crystal structure in complex with ecotin.
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DOI：
10.1371/journal.pone.0067962
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Thielens NM
中科院分区：
文献类型：
--
作者：
Gaboriaud C;Gupta RK;Martin L;Lacroix M;Serre L;Teillet F;Arlaud GJ;Rossi V;Thielens NM
已知甘露聚糖结合凝集素（MBL）、纤维胶凝蛋白和凝集素-11与三种同源模块蛋白酶MBL相关丝氨酸蛋白酶（MASP）相关。MASP-1和MASP-2的催化结构域的晶体结构已经解析，但MASP-3的相应结构域的结构仍然未知。最近发现MASP 1/3基因突变与以各种发育障碍为特征的罕见常染色体隐性3 MC（Mingarelli、Malpuech、Michels和Mingarvale）综合征之间的联系，揭示了MASP-3在早期发育过程中意想不到的重要作用。为了获得对MASP-3的酶性质和结构性质的初步了解，产生并表征了其丝氨酸蛋白酶（SP）结构域的重组形式。该结构域对荧光肽基-氨甲基香豆素底物的酰胺分解活性被证明比C1 r/C1 s/MASP家族的其他成员低得多。急诊大肠杆菌蛋白酶抑制剂ecotin与MASP-3和MASP-2的SP结构域结合，而与MASP-1、C1 r和C1 s没有检测到显著的相互作用。分离包含内皮素二聚体和两个MASP-3 SP结构域的四聚体复合物，并解析其晶体结构并将其精制至3.2 μ g。ecotin/MASP-3界面的分析允许更好地理解C1 r/C1 s/MASP蛋白酶家族成员对ecotin的不同反应性，并且MASP-3 SP结构域结构与其他胰蛋白酶样蛋白酶的结构域结构的比较产生了解释其体外酶原样性质的新假设。
Mannan-binding lectin (MBL), ficolins and collectin-11 are known to associate with three homologous modular proteases, the MBL-Associated Serine Proteases (MASPs). The crystal structures of the catalytic domains of MASP-1 and MASP-2 have been solved, but the structure of the corresponding domain of MASP-3 remains unknown. A link between mutations in the MASP1/3 gene and the rare autosomal recessive 3MC (Mingarelli, Malpuech, Michels and Carnevale,) syndrome, characterized by various developmental disorders, was discovered recently, revealing an unexpected important role of MASP-3 in early developmental processes. To gain a first insight into the enzymatic and structural properties of MASP-3, a recombinant form of its serine protease (SP) domain was produced and characterized. The amidolytic activity of this domain on fluorescent peptidyl-aminomethylcoumarin substrates was shown to be considerably lower than that of other members of the C1r/C1s/MASP family. The E. coli protease inhibitor ecotin bound to the SP domains of MASP-3 and MASP-2, whereas no significant interaction was detected with MASP-1, C1r and C1s. A tetrameric complex comprising an ecotin dimer and two MASP-3 SP domains was isolated and its crystal structure was solved and refined to 3.2 Å. Analysis of the ecotin/MASP-3 interfaces allows a better understanding of the differential reactivity of the C1r/C1s/MASP protease family members towards ecotin, and comparison of the MASP-3 SP domain structure with those of other trypsin-like proteases yields novel hypotheses accounting for its zymogen-like properties in vitro.
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影响因子：
5.7
作者：
Budayova-Spano, M;Grabarse, W;Gaboriaud, C
通讯作者：
Gaboriaud, C
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：
4.6
作者：
Di Cera, Enrico
通讯作者：
Di Cera, Enrico
DOI：
10.1107/s0907444998003254
发表时间：
1998-09-01
期刊：
ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY
影响因子：
--
作者：
Brunger, AT;Adams, PD;Warren, GL
通讯作者：
Warren, GL
影响因子：
11.4
作者：
Gaboriaud, C;Rossi, V;Fontecilla-Camps, JC
通讯作者：
Fontecilla-Camps, JC