Abstract
Human apurinic/apyrimidinic endonuclease-1 (APE-1) is essential for base excision repair and plays a major role in DNA repair and maintaining genomic stability. Cancer cells treated with conventional DNA-damaging agents develop resistance due in part to upregulation of enzymes involved in DNA repair. It is hypothesized that inhibiting DNA repair machinery should sensitize the cells to DNA-damaging agents. Previously, it has been shown that APE-1 is implicated in drug resistance and cancer progression. Therefore, APE-1 inhibitors are being sought after for their synergistic properties with various chemotherapeutics agents. Screening of several compound libraries and optimization of known inhibitors of APE-1 endonuclease activity have been accelerated by the use of high-throughput screening. Nevertheless, potential inhibitors must be tested in other counterscreens to validate their selectivity for APE-1. Here, we describe in-depth protocols for APE-1 purification and development of assays specific for APE-1 endonuclease activity.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Zawahir Z, Dayam R, Deng J, Pereira C, Neamati N (2009) Pharamacophore guided discovery of small-molecule human apurinic/apyrimidinic endonuclease 1 inhibitors. J Med Chem 52:20–32
Naidu MD, Mason JM, Pica RV, Fung H, Pena LA (2010) Radiation resistance in Glioma cells determined by DNA damage repair activity of APE-1/Ref-1. J Radiat Res 51:393–404
Lam W, Park SY, Leung CH, Cheng YC (2006) Apurinic/apyrimidinic endonuclease-1 protein level is associated with the cytotoxicity of l-configuration deoxycytidine analogs (troxacitabine and B-l-2′, 3′-dideoxy2′, 3′-didehydro-5-fluorocytidine) but not d-configuration deoxycytidine analogs (gemcitabine and B-d-arabinofuranosylcytosine). Mol Pharmacol 69:1607
Okagbare IP, Soper AS (2010) Polymer-based dense fluidic networks for high throughput screening with ultrasensitive fluorescence detection. Electrophoresis 31:3074–3082
Bapat A, Glass LS, Kuo M, Fishel ML, Long EC, Georgiadis MM, Kelley MR (2010) Novel small-molecule inhibitor of Apurinic/Apyrimidinic endonuclease 1 blocks proliferation and reduces viability of glioblastoma cells. J Pharmacol Toxicol Methods 334:988–998
Mohammed MZ, Vyjayanti VN, Laughton CA, Dekker LV, Fischer PM, Wilson DM 3rd, Abbotts R, Shah S, Patel PM, Hickson ID, Madhusudan S (2011) Development and evaluation of human AP endonuclease inhibitors in melanoma and glioma cell lines. Br J Cancer 104:653–66
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media New York
About this protocol
Cite this protocol
Esqueda, A., Mohammed, M.Z., Madhusudan, S., Neamati, N. (2012). Purification and Specific Assays for Measuring APE-1 Endonuclease Activity. In: Zheng, Y. (eds) Rational Drug Design. Methods in Molecular Biology, vol 928. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-008-3_13
Download citation
DOI: https://doi.org/10.1007/978-1-62703-008-3_13
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-007-6
Online ISBN: 978-1-62703-008-3
eBook Packages: Springer Protocols