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Proteases in Apoptosis: Pathways, Protocols and Translational Advances pp 143–202Cite as

Proteases in Apoptosis: Protocols and Methods

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Abstract

Proteases in apoptosis have evolved as major drug targets in the past few decades. Development in this direction has been brought about by efficient design and refinement of the various platforms of protease assays. These can be broadly categorized into general assays, that characterize kinetics and biochemistry of apoptotic proteases, and the more specific assays devoted to discern proteases involved in apoptosis. Together, these two approaches comprise a flawless two-pronged approach to understand the role of proteases in apoptosis and their therapeutic applications. This chapter lays down a comprehensive account of different experimental procedures spanning the use of in vitro purified proteases to those that monitor enzyme activity and its apoptotic effect in fixed or live cells. In this regard, fluorescence based platforms are the workhorse of fast, accurate, easy-to-use and high throughput screening amenable procedures. Therefore, they form the majority of techniques, among others, covered in this chapter. Apart from the popular methods currently in use, this chapter also provides a bird’s eye view of the future of the protease assays with special mention of protease activatable prodrugs and protease engineering.

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  1. Integrated Biophysics and Structural Biology Lab, Advanced Centre for Treatment, Research and Education in Cancer (ACTREC), Tata Memorial Centre, Navi Mumbai, Maharashtra, 410210, India

    Saujanya Acharya, Raja Reddy Kuppili, Lalith K. Chaganti & Kakoli Bose

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    Kakoli Bose

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Acharya, S., Kuppili, R.R., Chaganti, L.K., Bose, K. (2015). Proteases in Apoptosis: Protocols and Methods. In: Bose, K. (eds) Proteases in Apoptosis: Pathways, Protocols and Translational Advances. Springer, Cham. https://doi.org/10.1007/978-3-319-19497-4_5

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