Ultrasensitive internally quenched substrates of human cathepsin L
Ultrasensitive internally quenched substrates of human cathepsin L
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DOI：
10.1016/j.ab.2014.08.010
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发表时间：
2014-12-01
影响因子：
2.9
通讯作者：
Lesner, Adam
中科院分区：
文献类型：
--
作者：
Legowska, Monika;Wysocka, Magdalena;Lesner, Adam
合成了具有高特异性常数（k（cat）/K-M = 2.6 × 10（7）M-1 s（-1））的内淬灭的组织蛋白酶L（Cat L）底物ABZ-Bip-Arg-Ala-Gln-Tyr（3-NO2）-NH 2。所得化合物显示出对组织蛋白酶家族的其它成员（B、S、X、V、C、K、H、F、D和A）的高选择性。使用该底物发现Cat L在皮摩尔（pM）浓度下的活性。此外，确定了选择性Cat L抑制剂的存在将底物的蛋白水解抑制至不可检测的水平。将合成的化合物与健康细胞系和癌细胞系的细胞裂解物一起孵育表明Cat L活性存在显著差异。基于所获得的结果，它建议，这种基板可用于选择性监测的Cat L活性在生物系统中。(C)2014爱思唯尔公司All rights reserved.
Internally quenched cathepsin L (Cat L) substrate ABZ-Bip-Arg-Ala-Gln-Tyr(3-NO2)-NH2 with high specificity constant (k(cat)/K-M = 2.6 x 10(7) M-1 s(-1)) was synthesized. The resultant compound displayed high selectivity over other members of the cathepsin family (B, S, X, V, C, K, H, F, D, and A). Activity of Cat L at picomolar (pM) concentrations was found using this substrate. Moreover, it was established that the presence of the selective Cat L inhibitor suppressed the proteolysis of the substrate to a non-detectable level. Incubation of the synthesized compound with a cell lysate of healthy and cancer cell lines indicated significant differences in Cat L activity. Based on the obtained results, it is proposed that this substrate could be used for selective monitoring of Cat L activity in biological systems. (C) 2014 Elsevier Inc. All rights reserved.