Substrate specificity and inhibition of human kallikrein-related peptidase 3 (KLK3 or PSA) activated with sodium citrate and glycosaminoglycans.
Substrate specificity and inhibition of human kallikrein-related peptidase 3 (KLK3 or PSA) activated with sodium citrate and glycosaminoglycans.
复制标题
用柠檬酸钠和糖胺聚糖激活的人激肽释放酶相关肽酶 3（KLK3 或 PSA）的底物特异性和抑制。
DOI：
10.1016/j.abb.2010.03.022
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发表时间：
2010
影响因子：
3.9
通讯作者：
Juliano,Luiz
中科院分区：
文献类型：
--
作者：
Andrade,Douglas;Assis,DiegoM;Lima,AurelioResende;Oliveira,JulianaR;Araujo,MarianaS;Blaber,SachikoI;Blaber,Michael;Juliano,MariaA;Juliano,Luiz
我们报告了在糖胺聚糖（GAG）和柠檬酸钠存在下人重组KLK 3的酶性质和底物特异性。这种盐在前列腺中高度浓缩，并且在其存在下，KLK 3具有与胰凝乳蛋白酶相似的水解效率。与后一种肽酶相反，由柠檬酸钠激活的KLK 3有效地水解在P1位置含有R、H和P的底物。活化的KLK 3也在与人激肽释放酶KLK 1相同的位点切割来自人激肽原的缓激肽结构域的肽，但呈现低激肽原酶活性。血管紧张素I有几个被KLK 3水解的位点;然而，它仅在Y-I键处被切割（DRVY↓IHPFHL）。柠檬酸钠调制KLK 3构象观察到的苯丙氨酸和Dahans的固有荧光的改变。激活的KLK 3被Z-Pro-Prolinal可逆地抑制，并被邻菲咯啉竞争性地抑制。总之，这些是值得注意的观察结果，用于未来设计KLK 3的特异性非肽抑制剂和寻找天然底物。
We report the enzymatic properties and substrate specificity of human recombinant KLK3 in the presence of glycosaminoglycans (GAGs) and sodium citrate. This salt is highly concentrated in prostate and in its presence KLK3 had a similar hydrolytic efficiency as chymotrypsin. In contrast to the latter peptidase, KLK3 activated by sodium citrate efficiently hydrolyzed substrates containing R, H and P at the P1 position. Activated KLK3 also cleaved peptides derived from the bradykinin domain of human kininogen at the same sites as human kallikrein KLK1, but presented low kininogenase activity. Angiotensin I has several sites for hydrolysis by KLK3; however, it was cleaved only at the Y–I bond (DRVY↓IHPFHL). Sodium citrate modulated KLK3 conformation as observed by alterations to the intrinsic fluorescence of phenylalanines and tryptophans. Activated KLK3 was reversibly inhibited by Z-Pro-Prolinal and competitively inhibited by ortho-phenantroline. Together, these are noteworthy observations for the future design of specific non-peptide inhibitors of KLK3 and to find natural substrates.