Assays of matrix metalloproteinases (MMPs) activities: a review

Lombard, Carine; Saulnier, Joëlle; Wallach, Jean

doi:10.1016/j.biochi.2005.01.007

Cited by 93 publications

(63 citation statements)

References 71 publications

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“…Supernatants or lysates from treated endothelial cells are electrophoresed through the gel, followed by incubation in non-denaturing conditions to permit proteolysis. When zymogen gels are stained with a dye such as Coomassie blue, the hydrolyzed area appears transparent (reviewed in Lombard et al, 2005). The gel zymogen technique is inexpensive and provides information about the identities and relative levels of MMPs.…”

Section: Matrix Degradationmentioning

confidence: 99%

In vitro assays of angiogenesis for assessment of angiogenic and anti-angiogenic agents

Goodwin¹

2007

Microvascular Research

261

208

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Blood vessels, either in insufficient numbers or in excess, contribute to the pathogenesis of many diseases. Agents that stimulate angiogenesis can improve blood flow in patients with ischemic diseases, whereas anti-angiogenic agents are used to treat disorders ranging from macular degeneration to cancer. In this review I describe in vitro assays that can be used to assess the activity of agents that affect angiogenesis. Means of quantifying endothelial cell matrix degradation, migration, proliferation, apoptosis and morphogenesis are discussed, as are embryoid body, aortic ring and metatarsal assays of vessel outgrowth. Strengths and limitations of these techniques are also addressed.

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Section: Matrix Degradationmentioning

confidence: 99%

In vitro assays of angiogenesis for assessment of angiogenic and anti-angiogenic agents

Goodwin¹

2007

Microvascular Research

261

208

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“…In 2005, Lombard et al [6] published a very important review about MMPs activities assay, in which the assay methods were classified mainly by substrates, such as proteins, synthetic linear peptides and synthetic "mini-collagens". In order to summarize the further development of MMPs assay methods since 2005, such as in vivo zymography and activitybased profiling assay, we delivered this review.…”

Section: Introductionmentioning

confidence: 99%

Advances in assays of matrix metalloproteinases (MMPs) and their inhibitors

Cheng

Fang

2008

Journal of Enzyme Inhibition and Medicinal Chemistry

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Matrix metalloproteinases (MMPs) play an important role in many physiological and pathological processes. To assay the activities of MMPs is important in diagnosis and therapy of the MMPs associated diseases, such as neoplastic, rheumatic and cardiovascular diseases. Several assay systems have been developed, which include bioassay, zymography assay, immunoassay, fluorimetric assay, radio isotopic assay, phage-displayed assay, multiple-enzyme/multiple-reagent assay and activity-based profiling assay. The principle, application, advantage and disadvantage of these assays have been reviewed in this article.

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“…A continuous assay method, such as one that utilizes an increase in fluorescence upon hydrolysis, allows for rapid and convenient kinetic evaluation of proteases, both in solution and cell surface bound. For this reason, significant research efforts have focused on designing substrates for MMP family members (8). Sequence specificity studies for MMPs have been performed with peptides based on protein sequences surrounding MMP cleavage sites, and the results of these studies have been comprehensively reviewed (9,10).…”

Section: Development Of Peptide Substrates For Matrix Metalloproteinasesmentioning

confidence: 99%

“…fTHPs have subsequently provided insights into (a) the relative triple-helical peptidase activities of the various collagenolytic MMPs, (b) the collagen preferences of these MMPs, and (c) the relative roles of MMP domains and specific residues in efficient collagenolysis (50,52,53,57,58,(60)(61)(62)(63)(64)(65)(66). fTHPs have been noted as being particularly advantageous for dissecting the mechanism of collagenolysis (8).…”

Section: Triple-helicalmentioning

confidence: 99%

Using Fluorogenic Peptide Substrates to Assay Matrix Metalloproteinases

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Matrix Metalloproteinase Protocols

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A continuous assay method, such as one that utilizes an increase in fluorescence upon hydrolysis, allows for rapid and convenient kinetic evaluation of proteases. To better understand MMP behaviors and to aid in the design of MMP inhibitors, a variety of sequence specificity, phage display, and combinatorial chemistry studies have been performed. Results of these studies have been valuable for defining the differences in MMPs and for creating quenched fluorescent substrates that utilize fluorescence resonance energy transfer (FRET)/intramolecular fluorescence energy transfer (IFET). FRET triple-helical substrates have been constructed to examine the collagenolytic activity of MMP family members. The present chapter provides an overview of MMP and related FRET substrates and describes how to construct and utilize these substrates.

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Assays of matrix metalloproteinases (MMPs) activities: a review

Cited by 93 publications

References 71 publications

In vitro assays of angiogenesis for assessment of angiogenic and anti-angiogenic agents

In vitro assays of angiogenesis for assessment of angiogenic and anti-angiogenic agents

Advances in assays of matrix metalloproteinases (MMPs) and their inhibitors

Using Fluorogenic Peptide Substrates to Assay Matrix Metalloproteinases

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