Study on the Detection of MMP-2，-7，-9，and -12 Enzymatic Activity Using CEACAM1-Derived Fluorescent Peptide Substrate Site 84

Wen WAN; Yujia YE; Xiaona YANG; Lihong YANG; Huawei WANG; Ling DONG; Lixing CHEN; Zhaohui MENG

doi:10.12259/j.issn.2095-610X.S20250202

Volume 46 Issue 2

Feb. 2025

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Wen WAN, Yujia YE, Xiaona YANG, Lihong YANG, Huawei WANG, Ling DONG, Lixing CHEN, Zhaohui MENG. Study on the Detection of MMP-2，-7，-9，and -12 Enzymatic Activity Using CEACAM1-Derived Fluorescent Peptide Substrate Site 84[J]. Journal of Kunming Medical University, 2025, 46(2): 9-16. doi: 10.12259/j.issn.2095-610X.S20250202

Citation:

Wen WAN, Yujia YE, Xiaona YANG, Lihong YANG, Huawei WANG, Ling DONG, Lixing CHEN, Zhaohui MENG. Study on the Detection of MMP-2，-7，-9，and -12 Enzymatic Activity Using CEACAM1-Derived Fluorescent Peptide Substrate Site 84[J]. Journal of Kunming Medical University, 2025, 46(2): 9-16. doi: 10.12259/j.issn.2095-610X.S20250202

Citation:

Wen WAN, Yujia YE, Xiaona YANG, Lihong YANG, Huawei WANG, Ling DONG, Lixing CHEN, Zhaohui MENG. Study on the Detection of MMP-2，-7，-9，and -12 Enzymatic Activity Using CEACAM1-Derived Fluorescent Peptide Substrate Site 84[J]. Journal of Kunming Medical University, 2025, 46(2): 9-16. doi: 10.12259/j.issn.2095-610X.S20250202

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Study on the Detection of MMP-2，-7，-9，and -12 Enzymatic Activity Using CEACAM1-Derived Fluorescent Peptide Substrate Site 84

doi: 10.12259/j.issn.2095-610X.S20250202

1.
Department 1 of Cardiology，The First Affiliated Hospital of Kunming Medical University，Kunming Yunnan 650032
2.
Department 3 of Cardiology，The First Affiliated Hospital of Kunming Medical University，Kunming Yunnan 650032，China

Received Date: 2024-08-12
Available Online: 2025-01-14
Publish Date: 2025-02-18

Abstract

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 = 2565/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.
- Site 84,
- Fluorescent polypeptide substrate,
- Enzymatic activity,
- Gelatinase,
- CEACAM1-derived.

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