The efficient expression of human fibroblast collagenase in Escherichia coli and the discovery of flavonoid inhibitors.

Abstract

Human skin fibroblast collagenase also known as Matrix Metalloproteinase-1 (MMP-1) is a key enzyme in remodeling and degradation of extracellular matrix, and the inhibitors of human MMP-1 are effective drug candidates for the treatment of cancer. In this study, we report an improved method for high-level expression of soluble human MMP-1 catalytic domain (cd-MMP-1) in E.coli. The enzymatic activity is found maximum at pH 7.5 and temperature 40°C with a Km value of 13.02 µM. Effects of 17 structure-related flavonoids on MMP-1 activity are evaluated using a fluorescent assay, 6 inhibitors are identified with IC₅₀ < 10 µM. Fisetin is the most active agent with an IC₅₀ value of 1.35 µM and is identified as a mixed type inhibitor. Our improved soluble cd-MMP-1 expression method provides a basis for inhibitors identification and may be beneficial to discover novel anti-cancer agent targeting human MMP-1.

• Authors: Weiqiang Lu, Junsheng Zhu, Shien Zou, Xi Li, Jin Huang
• Journal: Journal of enzyme inhibition and medicinal chemistry (J Enzyme Inhib Med Chem) Vol. 28 Issue 4 Pg. 741-6 (Aug 2013) ISSN: 1475-6374 [Electronic] England
• PMID: 22524676 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Enzyme Inhibitors
• Flavonoids
• Matrix Metalloproteinase 8

Topics

• Cloning, Molecular
• Dose-Response Relationship, Drug
• Drug Discovery
• Enzyme Activation (drug effects)
• Enzyme Inhibitors (chemical synthesis, chemistry, pharmacology)
• Escherichia coli (genetics, metabolism)
• Flavonoids (chemical synthesis, chemistry, pharmacology)
• Gene Expression Regulation, Enzymologic (drug effects, genetics)
• Humans
• Matrix Metalloproteinase 8 (genetics, metabolism)
• Molecular Structure
• Structure-Activity Relationship