Rozman-Pungercar J, Kopitar-Jerala N, Bogyo M, Turk D, Vasiljeva O, Stefe I, Vandenabeele P, Brömme D, Puizdar V, Fonović M, Trstenjak-Prebanda M, Dolenc I, Turk V, Turk B
Department of Biochemistry and Molecular Biology, J. Stefan Institute, Jamova 39, SI-1000 Ljubljana, Slovenia.
Cell Death Differ. 2003 Aug;10(8):881-8. doi: 10.1038/sj.cdd.4401247.
We report here that a number of commonly used small peptide caspase inhibitors consisting of a caspase recognition sequence linked to chloromethylketone, fluoromethylketone or aldehyde reactive group efficiently inhibit other cysteine proteases than caspases. The in vitro studies included cathepsins B, H, L, S, K, F, V, X and C, papain and legumain. Z-DEVD-cmk was shown to be the preferred irreversible inhibitor of most of the cathepsins in vitro, followed by Z-DEVD-fmk, Ac-YVAD-cmk, Z-YVAD-fmk and Z-VAD-fmk. Inactivation of legumain by all the inhibitors investigated was moderate, whereas cathepsins H and C were poorly inhibited or not inhibited at all. Inhibition by aldehydes was not very potent. All the three fluoromethylketones efficiently inhibited cathepsins in Jurkat and human embryonic kidney 293 cells at concentrations of 100 microM. Furthermore, they completely inhibited cathepsins B and X activity in tissue extracts at concentrations as low as 1 microM. These results suggest that data based on the use of these inhibitors should be taken with caution and that other proteases may be implicated in the processes previously ascribed solely to caspases.
我们在此报告，一些常用的小肽半胱天冬酶抑制剂，由与氯甲基酮、氟甲基酮或醛反应基团相连的半胱天冬酶识别序列组成，能有效抑制除半胱天冬酶之外的其他半胱氨酸蛋白酶。体外研究包括组织蛋白酶B、H、L、S、K、F、V、X和C、木瓜蛋白酶和天冬酰胺酶。在体外，Z-DEVD-cmk被证明是大多数组织蛋白酶的首选不可逆抑制剂，其次是Z-DEVD-fmk、Ac-YVAD-cmk、Z-YVAD-fmk和Z-VAD-fmk。所研究的所有抑制剂对天冬酰胺酶的失活作用中等，而组织蛋白酶H和C受抑制程度较差或根本未受抑制。醛类的抑制作用不太有效。所有三种氟甲基酮在100微摩尔浓度下能有效抑制Jurkat细胞和人胚肾293细胞中的组织蛋白酶。此外，它们在低至1微摩尔的浓度下就能完全抑制组织提取物中组织蛋白酶B和X的活性。这些结果表明，基于使用这些抑制剂得出的数据应谨慎对待，并且其他蛋白酶可能参与了以前仅归因于半胱天冬酶的过程。
