Convenient fluorometric assay for matrix metalloproteinase activity and its application in biological media
Convenient fluorometric assay for matrix metalloproteinase activity and its application in biological media
复制标题
DOI：
10.1016/0014-5793(96)00665-5
复制
发表时间：
1996-07-22
期刊：
影响因子：
3.5
通讯作者：
Koppele, JMT
中科院分区：
文献类型：
--
作者：
Beekman, B;Drijfhout, JW;Koppele, JMT
基质金属蛋白酶（MMPs）参与生理组织重塑和病理条件，如肿瘤转移和关节破坏。到目前为止，没有方便和敏感的MMP活性测定在粗介质，如滑液已可用。因此，合成了高度可溶的荧光底物TNO 211（Dabcyl-Gaba-Pro-Gln-Gly-Leu-Glu（EDANS）-Ala-Lys-NH 2），其含有MMP可裂解的Gly-Leu键和EDANS/Dabcyl作为荧光团/测序剂组合，并表征为MMP特异性底物。结果表明，TNO 211荧光法检测RA和OA患者及对照组的滑膜液（SF）和内皮细胞培养液中MMP活性灵敏度高，SF中MMP活性按C < OA < RA的顺序显著升高，因此，在RA研究中经常使用OA样本作为对照是值得商榷的。
Matrix metalloproteinases (MMPs) are involved in physiological tissue remodeling and pathological conditions like tumour metastasis and joint destruction. Until now, no convenient and sensitive MMP-activity assay in crude media like synovial fluid has been available. Therefore, the highly soluble fluorogenic substrate TNO211 (Dabcyl-Gaba-Pro-Gln-Gly-Leu-Glu(EDANS)-Ala-Lys-NH2), containing the MMP cleavable Gly-Leu bond and EDANS/Dabcyl as fluorophore/quencer combination, was synthesized and characterized as an MMP specific substrate. We show that the fluorogenic assay using TNO211 is sensitive and can detect MMP activity in culture medium from endothelial cells and untreated synovial fluid (SF) from RA and OA patients, and control subjects, MMP activity in SF significantly increased in the order C < OA < RA, thus the frequent use of OA samples as control in studies on RA is debatable.