Analysis of enzymatic activity of matrix metalloproteinase (MMP) by collagen zymography in Melanoma

Vijay Walia, Yardena Samuels*

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapter

3 Citations (Scopus)

Abstract

Protein zymography is the most commonly used technique to study the enzymatic activity of matrix metalloproteinases (MMPs) and their inhibitors. MMPs are proteolytic enzymes that promote extracellular matrix degradation. MMPs are frequently mutated in malignant melanomas as well as other cancers and are linked to increasing incidence of tumor metastasis. Substrate zymography characterizes MMP activity by their ability to degrade preferred substrates. Here we describe the collagen zymography technique to measure the active or latent form of MMPs using MMP-8 as an example, which is a frequently mutated MMP family member in malignant melanomas. The same technique can be used with the modification of substrate to detect metalloproteinase activity of other MMPs. Both wild-type and mutated forms of MMPs can be analyzed using a single gel using this method.

Original languageEnglish
Title of host publicationProteases and Cancer
EditorsS. Cal, A. Obaya
PublisherHumana Press
Pages97-106
Number of pages10
ISBN (Electronic)978-1-4939-7595-2
ISBN (Print)978-1-4939-7594-5
DOIs
Publication statusPublished Online - 10 Jan 2018

Publication series

SeriesMethods in Molecular Biology
Volume1731
ISSN1064-3745

Bibliographical note

Y.S. is supported by the ERC (StG-335377), by the MRA Young Investigator Award #402024 by the Henry Chanoch Krenter Institute for Biomedical Imaging and Genomics, the estate of Alice Schwarz-Gardos, the estate of John Hunter, the Knell Family, the Peter and Patricia Gruber Award, and the Hamburger Family.

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

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