Measurement of activity of collagenolytic MMP and inhibitors of MMPs using radiolabeled collagen substrate.

Abstract:

:This protocol describes how to purify and radiolabel collagen for use as a substrate to assay collagenolytic members of the matrix metalloproteinases (MMPs). This assay measures enzymes that specifically cleave native triple helical collagen. After incubation of the MMP enzyme with the collagen substrate at 37 degrees C, undigested collagen is removed by centrifugation. Radiolabeled cleaved fragments remain in the supernatant, which is then counted in a scintillation counter; a linear increase in the release of radiolabeled collagen fragments occurs with enzyme level and time. Methods are included for the activation of the proenzyme forms of these MMPs and the assay can also be adapted to measure inhibitors of the collagenolytic MMPs. This assay can be completed in 18 h.

journal_name

Nat Protoc

journal_title

Nature protocols

authors

Cawston TE

doi

10.1038/nprot.2008.244

subject

Has Abstract

pub_date

2009-01-01 00:00:00

pages

286-90

issue

3

eissn

1754-2189

issn

1750-2799

pii

nprot.2008.244

journal_volume

4

pub_type

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