Characterization of Mg2+ Binding to the DNA Repair Protein Apurinic/Apyrimidic Endonuclease 1 via Solid-State 25Mg NMR Spectroscopy

Autor:	Robert W. Heck, D. M. Wilson, Andrew S. Lipton, D. R. McNeill, S. Primak, Paul D. Ellis
Rok vydání:	2008
Předmět:	biology Stereochemistry Chemistry General Chemistry Nuclear magnetic resonance spectroscopy Base excision repair Biochemistry Catalysis AP endonuclease Endonuclease chemistry.chemical_compound Colloid and Surface Chemistry DNA Repair Protein biology.protein AP site Binding site DNA
Zdroj:	Journal of the American Chemical Society. 130:9332-9341
ISSN:	1520-5126 0002-7863
DOI:	10.1021/ja0776881
Popis:	Apurinic/apyrimidinic endonuclease 1 (APE1), a member of the divalent cation-dependent phosphoesterase superfamily of proteins that retain the conserved four-layered α/β-sandwich structural core, is an essential protein that functions as part of base excision repair to remove mutagenic and cytotoxic abasic sites from DNA. Using low-temperature solid-state 25Mg NMR spectroscopy and various mutants of APE1, we demonstrate that Mg2+ binds to APE1 and a functional APE1−substrate DNA complex with an overall stoichiometry of one Mg2+ per mole of APE1 as predicted by the X-ray work of Tainer and co-workers (Mol, C. D.; Kuo, C. F.; Thayer, M. M.; Cunningham, R. P.; Tainer, J. A. Nature 1995, 374, 381−386). However, the NMR spectra show that the single Mg2+ site is disordered. We discuss the probable reasons for the disorder at the Mg2+ binding site. The most likely source of this disorder is arrangement of the protein−ligands about the Mg2+ (cis and trans isomers). The existence of these isomers reinforces the no...
Databáze:	OpenAIRE
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