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Zymography as a Research Tool in the Study of Matrix Metalloproteinase Inhibitors

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Zymography

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1626))

Abstract

Matrix metalloproteinases (MMPs) are proteolytic enzymes that degrade various components of the extracellular matrix (ECM) and play a role in tissue remodeling. Changes in MMPs have been observed in cancer, connective tissue disorders, and vascular disease, and both endogenous tissue inhibitors of MMPs (TIMPs) and synthetic MMP inhibitors (MMPIs) have been evaluated as modulators of MMP activity in various biological systems. Zymography is a simple technique that is commonly used to assess MMP activity and the efficacy of MMPIs. Also, reverse zymography is a modified technique to study the activity of endogenous TIMPs. However, problems are often encountered during the zymography procedure, which could interfere with accurate assessment of MMP activity in control specimens, and thus make it difficult to determine the pathological changes in MMPs and their responsiveness to MMPIs. Simplified protocols for preparation of experimental solutions, tissue preparation, regular and reverse zymography procedures, and zymogram analysis are presented. Additional helpful tips to troubleshoot problems in the zymography technique and to enhance the quality of the zymograms should make it more feasible to determine the changes in MMPs and assess the efficacy of MMPIs in modulating MMP activity in various biological systems and pathological conditions.

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Abbreviations

A/C:

Acrylamide/bis-acrylamide

ECM:

Extracellular matrix

MMP:

Matrix metalloproteinase

MMPI:

MMP inhibitor

MT-MMP:

Membrane-type-MMP

RUPP:

Reduction in uterine perfusion pressure

TIMP:

Tissue inhibitor of MMP

Zn2+ :

Zinc ion

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Acknowledgments

This work was supported by grants from National Heart, Lung, and Blood Institute (HL-65998, HL-98724, HL-111775). Dr. Zongli Ren was a visiting scholar from the Department of Cardiovascular Surgery, Renmin Hospital of Wuhan University, Wuhan 430060, Hubei, PR China, and a recipient of scholarship from the China Scholarship Council. Dr. Juanjuan Chen was a visiting scholar from the Department of Obstetrics & Gynecology, The Third Affiliated Hospital of Guangzhou Medical University, Guangzhou, China 510150.

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Authors and Affiliations

  1. Vascular Surgery Research Laboratory, Division of Vascular and Endovascular Surgery, Brigham and Women’s Hospital and Harvard Medical School, 75 Francis Street, Boston, MA, 02115, USA

    Zongli Ren, Juanjuan Chen & Raouf A. Khalil M.D., Ph.D.

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  1. Zongli Ren
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  2. Juanjuan Chen
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  3. Raouf A. Khalil M.D., Ph.D.
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Correspondence to Raouf A. Khalil M.D., Ph.D. .

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  1. Centre for Environmental, Biology and Chemistry Research, Facultad de Ciencias y Tecnología, University of Carabobo, Valencia, Venezuela

    Jeff Wilkesman

  2. Centre for Environmental, Biology and Chemistry Research Facultad de Ciencias y Tecnología, University of Carabobo, Valencia, Venezuela

    Liliana Kurz

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Ren, Z., Chen, J., Khalil, R.A. (2017). Zymography as a Research Tool in the Study of Matrix Metalloproteinase Inhibitors. In: Wilkesman, J., Kurz, L. (eds) Zymography. Methods in Molecular Biology, vol 1626. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7111-4_8

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  • DOI: https://doi.org/10.1007/978-1-4939-7111-4_8

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