昆明医科大学学报2025，Vol.46Issue(2)：9-16,8.DOI:10.12259/j.issn.2095-610X.S20250202
CEACAM1源性荧光多肽底物Site 84对MMP-2、-7、-9、-12酶活性检测的研究
Study on the Detection of MMP-2,-7,-9,and-12 Enzymatic Activity Using CEACAM1-Derived Fluorescent Peptide Substrate Site 84
摘要
Abstract
Objective To explore the the detection of MMP-2,-7,-9,and-12 enzymatic activity using the CEACAM1-derived fluorescent peptide substrate Site 84,investigating the application of substrate Site 84 to distinguishing between MMP-2 and MMP-9 in the gelatinase spectrum of MMPs.Methods The fluorescent enzymatic method was employed to observe the detection of MMP-2,-7,-9,and-12 enzymatic activity using substrate Site 84;further observations were made on the sensitivity and specificity of substrate Site 84 to enzymatic activity of MMP-2 and MMP-9 within the gelatinase spectrum;the kinetic parameters(Km and Kcat)of the enzymatic reaction between substrate Site 84 and MMP-2 were obtained.Results Using Site 84 as a substrate,enzymic kinetics curves for MMP-12,-7,-2 were obtained,but no enzymatic activity curve for MMP-9 was observed.Furthermore,Site 84 specifically detected the enzymatic activity of MMP-2 within the gelatinase spectrum,capable of detecting low concentration(0.6 μM)of MMP-2 enzymatic activity,but no obvious enzymatic reaction was observed for high concentration(6 μM)of MMP-9;the kinetics parameters for the enzymatic reaction between Site 84 and MMP-2 were Km=315 μM,Kcat/Km=2 565/MS.Conclusion The CEACAM1-derived substrate Site 84 serves as a novel fluorescent peptide substrate,enabling the acquisition of enzymatic activity curves for MMP-12,-7 and-2,and specifically detecting the enzymatic activity of MMP-2 within the MMP gelatinase spectrum.关键词
Site 84/荧光多肽底物/酶活性/明胶酶/CEACAM1源性Key words
Site 84/Fluorescent polypeptide substrate/Enzymatic activity/Gelatinase/CEACAM1-derived分类
医药卫生引用本文复制引用
万雯,叶雨佳,杨晓娜,杨理宏,王华炜,董玲,陈丽星,孟照辉..CEACAM1源性荧光多肽底物Site 84对MMP-2、-7、-9、-12酶活性检测的研究[J].昆明医科大学学报,2025,46(2):9-16,8.基金项目
国家自然科学基金(82360077) （82360077）
云南省基础研究计划昆医联合专项-面上项目(202201AY070001-082) （202201AY070001-082）
云南省基础研究计划青年项目(202201AU070063) （202201AU070063）
昆明医科大学第一附属医院"535"高层次人才(2024535Q03) （2024535Q03）
