The enzymes Matrix metalloproteinases (MMPs) and A Disintegrin and Metalloproteinases (ADAMs) are two related protease families which play key roles in physiological processes. MMPs are involved in processes such as development, angiogenesis and tissue remodeling, whilst ADAMs mediate various cellular behaviour. Many useful methodologies currently exist to study metalloproteinases but none simultaneously allow for direct, non-invasive, multiplex, real-time measurements of specific protease activity.
Proteolytic Activity Matrix Analysis (PrAMA) is an integrated experimental measurement and mathematical analysis framework, developed by Douglas A. Lauffenburger et al., Massachusetts Institute of Technology, in an attempt to address this issue.
This method uses panels of FRET-substrates to infer a dynamic, quantitative and specific profile of MMP and ADAM proteolytic activities and by utilising predetermined signatures, from purified enzymes, it is possible to deconvolute signals from complex protease mixtures to ascertain specific protease activity. PrAMA appears to be able to distinguish closely related enzymes with a high accuracy and can do so even when there is unknown background proteolytic activity present.
Lauffenburger et al. believe that this has the potential to be used in areas ranging from systems biology to in vitro inhibitor screening as well as other protease families.
Find out more about PrAMA by reading this HOT article, which is part of a forthcoming Integrative Biology themed issue in honour of Mina J. Bissell. Go on, it’s free for 4 weeks!
Proteolytic Activity Matrix Analysis (PrAMA) for simultaneous determination of multiple protease activities
Miles A. Miller, Layla Barkal, Karen Jeng, Andreas Herrlich, Marcia Moss, Linda G. Griffith and Douglas A. Lauffenburger
Integr. Biol., 2011, Advance Article
This article was published as part of a themed issue in honor of Mina J. Bissell.