School of Biological Sciences and Technology, Chonnam National University, 77 Yongbong-ro, Buk-gu, Gwangju 500-757, Republic of Korea.
Enzyme Microb Technol. 2013 Jan 10;52(1):26-31. doi: 10.1016/j.enzmictec.2012.10.001. Epub 2012 Oct 13.
The human fibroblast collagenase catalytic domain (MMP1ca) that is considered a prototype for all interstitial collagenase and plays an important role in the turnover of collagen fibrils in the matrix was expressed as an inclusion body in the Escherichia coli. The purified enzyme displayed activity with substrate Dnp-Pro-Leu-Ala-Leu-Trp-Ala-Arg-OH with a K(m) value of 26.61±1.42 μM. The inhibition activity of the nine flavonoid compounds and gallic acid against MMP1ca was examined. Among the compounds tested, the IC(50) of seven flavonoid compounds were determined and ranged from 14.13 to 339.21 μM. Epigallocatechin gallate (EGCG) showed the highest inhibition toward MMP1ca with IC(50) values of 14.13±0.49 μM. EGCG showed a competitive inhibition pattern with a K(i) value of 10.47±0.51 μM. The free binding energy of EGCG against MMP1ca was -13.07 kcal mol(-1), which was calculated by using Autodock 3.0.5 software and showed numerous hydrophobic and hydrogen bond interactions. The galloyl group of EGCG, gallocatechin gallate and epicatechin gallate was determined to be important for inhibitory activity against MMP1ca.
人成纤维细胞胶原酶催化结构域（MMP1ca）被认为是所有间质胶原酶的原型，在基质中胶原纤维的转化中发挥重要作用，在大肠杆菌中作为包涵体表达。纯化的酶对底物 Dnp-Pro-Leu-Ala-Leu-Trp-Ala-Arg-OH 显示出活性，其 K(m)值为 26.61±1.42 μM。考察了 9 种黄酮类化合物和没食子酸对 MMP1ca 的抑制活性。在所测试的化合物中，测定了 7 种黄酮类化合物的 IC（50），范围为 14.13-339.21 μM。表没食子儿茶素没食子酸酯（EGCG）对 MMP1ca 的抑制作用最强，IC（50）值为 14.13±0.49 μM。EGCG 表现出竞争性抑制模式，K(i)值为 10.47±0.51 μM。使用 Autodock 3.0.5 软件计算得出，EGCG 与 MMP1ca 的自由结合能为-13.07 kcal mol（-1），显示出许多疏水和氢键相互作用。EGCG、没食子儿茶素没食子酸酯和表儿茶素没食子酸酯的没食子酰基被确定为抑制 MMP1ca 活性的重要基团。
