Kwon Chang Woo, Yang Hee, Yeo SuBin, Park Kyung-Min, Jeong Ae Jin, Lee Ki Won, Ye Sang-Kyu, Chang Pahn-Shick
a Department of Agricultural Biotechnology , Seoul National University , Seoul , Republic of Korea.
b Department of Pharmacology and Biomedical Sciences , Seoul National University College of Medicine , Seoul , Republic of Korea.
J Enzyme Inhib Med Chem. 2018 Dec;33(1):657-664. doi: 10.1080/14756366.2018.1444609.
Cathepsin L of cancer cells has been shown to play an important role in degradation of extracellular matrix for metastasis. In order to reduce cell invasion, cathepsin L propeptide-like proteins which are classified as the I29 family in the MEROPS peptidase database were characterized from Calotropis procera R. Br., rich in cysteine protease. Of 19 candidates, the cloned and expressed recombinant SnuCalCp03-propeptide (rSnuCalCp03-propeptide) showed a low nanomolar K value of 2.3 ± 0.2 nM against cathepsin L. A significant inhibition of tumor cell invasion was observed with H1975, HT29, MDA-BM-231, PANC1, and PC3 with a 76, 67, 67, 63, and 79% reduction, respectively, in invasion observed in the presence of 400 nM of the rSnuCalCp03-propeptide. In addition, thermal and pH study showed rSnuCalCp03-propeptide consisting of secondary structures was stable at a broad range of temperatures (30-70 °C) and pH (2-10, except for 5 which is close to the isoelectric point of 5.2).
癌细胞中的组织蛋白酶L已被证明在转移过程中细胞外基质的降解中起重要作用。为了减少细胞侵袭，从富含半胱氨酸蛋白酶的牛角瓜（Calotropis procera R. Br.）中鉴定出在MEROPS肽酶数据库中被归类为I29家族的组织蛋白酶L前肽样蛋白。在19个候选蛋白中，克隆并表达的重组SnuCalCp03前肽（rSnuCalCp03 - 前肽）对组织蛋白酶L显示出2.3±0.2 nM的低纳摩尔K值。在400 nM的rSnuCalCp03 - 前肽存在下，观察到H1975、HT29、MDA - BM - 231、PANC1和PC3肿瘤细胞侵袭分别显著抑制，侵袭减少了76%、67%、67%、63%和79%。此外，热稳定性和pH研究表明，由二级结构组成的rSnuCalCp03 - 前肽在广泛的温度范围（30 - 70°C）和pH范围（2 - 10，除了接近5.2等电点的5）内是稳定的。
