Advances in the expression and purification of human PARP1: A user-friendly protocol
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https://hdl.handle.net/10037/33215Date
2023-07-05Type
Journal articleTidsskriftartikkel
Peer reviewed
Abstract
The PARP1 (Poly (ADP-ribose) polymerase 1) enzyme is essential for single and double-strand break repair in humans. Alterations affecting PARP1 activity have severe consequences for human health and are associated with pathologies like cancer, and metabolic and neurodegenerative disorders. Here, we have developed a fast and easy procedure for the expression and purification of PARP1. Biologically active protein was purified to an apparent purity > 95%, with only two purification steps. A thermostability analysis revealed that PARP1 possessed improved stability in 50 mM Tris-HCl pH 8.0 (Tm = 44.2 ± 0.3 °C), thus this buffer was used throughout the whole purification procedure. The protein was shown to bind to DNA and has no inhibitor molecules bound to the active site. Finally, the yield of the purified PARP1 protein is sufficient for both biochemical, biophysical and structural analysis. The new protocol provides a fast and simple purification procedure while producing similar protein quantities to what has been described previously.
Publisher
ElsevierCitation
Conceição, Salgueiro, Ribeiro, Raposo, Moe. Advances in the expression and purification of human PARP1: A user-friendly protocol. Protein Expression and Purification. 2023;211Metadata
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