Abstract
Peptide array-based in situ fluorescence assay is a reliable and efficient technique for high-throughput profiling and localization of enzyme activity. Here, peptide array is fabricated by spotting five specific MMPs (MMP-2, MMP-3, MMP-7, MMP-9, and MMP-14) peptide substrates containing FAM/Dabcyl fluorescent resonance energy transfer (FRET) pair on the surface of cell monolayers or tissue sections. MMP activities are determined in situ by the fluorescence intensity of stained cells/tissues due to the cellular internalization of hydrolyzed peptide fragments with FAM moieties. Identification of MMP expression patterns of cells, highly sensitive determination of MMP activities in cell monolayer (as low as hundreds of cells per square centimeter), and evaluation of inhibition potencies of six compounds toward five MMPs are achieved by this method. Five MMP activities in the localized parts of 32 thyroid tissues are also well profiled without separation or extraction procedures.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Nagase H, Woessner JF (1999) Matrix metalloproteinases. J Biol Chem 274:21491–21494
Malemud CJ (2006) Matrix metalloproteinases (MMPs) in health and disease: an overview. Front Biosci 11:1696–1701
Deryugina EI, Quigley JP (2006) Matrix metalloproteinases and tumor metastasis. Cancer Metastasis Rev 25:9–34
Köhrmann A, Kammerer U, Kapp M et al (2009) Expression of matrix metalloproteinases (MMPs) in primary human breast cancer and breast cancer cell lines: new findings and review of the literature. BMC Cancer 9:188
Baum O, Hlushchuk R, Forster A et al (2007) Increased invasive potential and up-regulation of MMP-2 in MDA-MB-231 breast cancer cells expressing the beta3 integrin subunit. Int J Oncol 30:325–332
Barbosa F Jr, Gerlach F, Tanus-Santos JE (2006) Matrix metalloproteinase-9 activity in plasma correlates with plasma and whole blood lead concentrations. Basic Clin Pharmacol Toxicol 98:559–564
Nie Y, Zhang P, Wang H et al (2017) Ultrasensitive electrochemiluminescence biosensing platform for detection of multiple types of biomarkers toward identical cancer on a single interface. Anal Chem 89:12821–12827
Cao S, Li Z, Zhao J et al (2018) Rational engineering a multichannel upconversion sensor for multiplex detection of matrix metalloproteinase activities. ACS Sens 3:1522–1530
Mook OR, Van Overbeek C, Ackema EG et al (2003) In situ localization of gelatinolytic activity in the extracellular matrix of metastases of colon cancer in rat liver using quenched fluorogenic DQ-gelatin. J Histochem Cytochem 51:821–829
Galis ZS, Sukhova GK, Libby P (1995) Microscopic localization of active proteases by in situ zymography: detection of matrix metalloproteinase activity in vascular tissue. FASEB J 9:974–980
Wang X, Qin L, Zhou M et al (2018) Nanozyme sensor arrays for detecting versatile analytes from small molecules to proteins and cells. Anal Chem 90:11696–11702
Qin L, Wang X, Liu Y et al (2018) 2D-metal-organic-framework-nanozyme sensor arrays for probing phosphates and their enzymatic hydrolysis. Anal Chem 90:9983–9989
Wang Y, Li H, Xu D (2016) Aptamers-based sandwich assay for silver-enhanced fluorescence multiplex detection. Anal Chim Acta 905:149–155
Qi H, Wang F, Petrenko VA et al (2014) Peptide microarray with ligands at high density based on symmetrical carrier landscape phage for detection of cellulase. Anal Chem 86:5844–5850
Sievers S, Cretich M, Gagni P et al (2017) Performance of a polymer coated silicon microarray for simultaneous detection of food allergen-specific IgE and IgG4. Clin Exp Allergy 47:1057–1068
Cabezas MD, Mirkin CA, Mrksich M (2017) Nanopatterned extracellular matrices enable cell-based assays with a mass spectrometric readout. Nano Lett 17:1373–1377
Li H, Brewer G, Ongo G et al (2017) Immunohistochemistry microarrays. Anal Chem 89:8620–8625
Lei Z, Zhang H, Wang Y et al (2017) Peptide microarray-based metal enhanced fluorescence assay for multiple profiling of matrix metalloproteinases activities. Anal Chem 89:6749–6757
Egeblad M, Werb Z (2002) New functions for the matrix metalloproteinases in cancer progression. Nat Rev Cancer 2:161
Hong Y, Ku M, Heo D et al (2014) Molecular recognition of proteolytic activity in metastatic cancer cells using fluorogenic gold nanoprobes. Biosens Bioelectron 57:171–178
Wang Y, Lin T, Zhang W et al (2015) A prodrug-type, MMP-2-targeting nanoprobe for tumor detection and imaging. Theranostics 5:787–795
Chung TW, Moon SK, Chang YC et al (2004) Novel and therapeutic effect of caffeic acid and caffeic acid phenyl ester on hepatocarcinoma cells: complete regression of hepatoma growth and metastasis by dual mechanism. FASEB J 18:1670–1681
Ende C, Gebhardt R (2004) Inhibition of matrix metalloproteinase-2 and -9 activities by selected flavonoids. Planta Med 70:1006–1008
Phromnoi K, Yodkeeree S, Anuchapreeda S et al (2009) Inhibition of MMP-3 activity and invasion of the MDA-MB-231 human invasive breast carcinoma cell line by bioflavonoids. Acta Pharmacol Sin 30:1169–1176
Moroy G, Bourguet E, Decarme M et al (2011) Inhibition of human leukocyte elastase, plasmin and matrix metalloproteinases by oleic acid and oleoyl-galardin derivative(s). Biochem Pharmacol 81:626–635
Maeta H, Ohgi S, Terada T (2001) Protein expression of matrix metalloproteinases 2 and 9 and tissue inhibitors of metalloproteinase 1 and 2 in papillary thyroid carcinomas. Virchows Arch 438:121–128
Acknowledgments
This work was supported by National Natural Science Foundation of China (grant no. 21775145).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Lei, Z., Wang, Z. (2023). Peptide Array-Based In Situ Fluorescence Assay for Profiling Multiple Matrix Metalloproteinase Activities. In: Cretich, M., Gori, A. (eds) Peptide Microarrays. Methods in Molecular Biology, vol 2578. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2732-7_12
Download citation
DOI: https://doi.org/10.1007/978-1-0716-2732-7_12
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-2731-0
Online ISBN: 978-1-0716-2732-7
eBook Packages: Springer Protocols