Study on the Detection of MMP-2,-7,-9,and-12 Enzymatic Activity Using CEACAM1-Derived Fluorescent Peptide Substrate Site 84
10.12259/j.issn.2095-610X.S20250202
- VernacularTitle: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
- Author:
Wen WAN
1
;
Yujia YE
;
Xiaona YANG
;
Lihong YANG
;
Huawei WANG
;
Ling DONG
;
Lixing CHEN
;
Zhaohui MENG
Author Information
1. 昆明医科大学第一附属医院心脏内一科,云南 昆明 650032
- Keywords:
Site 84;
Fluorescent polypeptide substrate;
Enzymatic activity;
Gelatinase;
CEACAM1-derived
- From:
Journal of Kunming Medical University
2025;46(2):9-16
- CountryChina
- Language:Chinese
-
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.
