Abstract
A general method to express and purify full-length human poly(ADP-ribose) polymerase-1 (PARP-1), individual PARP-1 domains, and groups of PARP-1 domains from Escherichia coli cells is described. The procedure allows for robust production of highly pure PARP-1 that is free of DNA contamination and well-suited for biochemical experiments and for structural and biophysical analysis. Two biochemical assays for monitoring PARP-1 automodification activity are presented that can be used to evaluate purified PARP-1, combinations of PARP-1 domains, or PARP-1 mutants.
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Langelier, MF., Planck, J.L., Servent, K.M., Pascal, J.M. (2011). Purification of Human PARP-1 and PARP-1 Domains from Escherichia coli for Structural and Biochemical Analysis. In: Tulin, A. (eds) Poly(ADP-ribose) Polymerase. Methods in Molecular Biology, vol 780. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-270-0_13
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DOI: https://doi.org/10.1007/978-1-61779-270-0_13
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-61779-269-4
Online ISBN: 978-1-61779-270-0
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