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A “moving metal mechanism” for substrate cleavage by the DNA repair endonuclease APE‐1
Author(s) -
Oezguen Numan,
Schein Catherine H.,
Peddi Srinivasa R.,
Power Trevor D.,
Izumi Tadahide,
Braun Werner
Publication year - 2007
Publication title -
proteins: structure, function, and bioinformatics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.699
H-Index - 191
eISSN - 1097-0134
pISSN - 0887-3585
DOI - 10.1002/prot.21397
Subject(s) - ap site , endonuclease , cleavage (geology) , dna , metal , chemistry , ap endonuclease , ion , base excision repair , molecular dynamics , metal ions in aqueous solution , crystallography , dna ligase , bond cleavage , stereochemistry , dna damage , biophysics , materials science , biochemistry , computational chemistry , biology , catalysis , organic chemistry , fracture (geology) , composite material
Apurinic/apyrimidinic endonuclease (APE‐1) is essential for base excision repair (BER) of damaged DNA. Here molecular dynamics (MD) simulations of APE1 complexed with cleaved and uncleaved damaged DNA were used to determine the role and position of the metal ion(s) in the active site before and after DNA cleavage. The simulations started from an energy minimized wild‐type structure of the metal‐free APE1/damaged‐DNA complex (1DE8). A grid search with one Mg 2+ ion located two low energy clusters of Mg 2+ consistent with the experimentally determined metal ion positions. At the start of the longer MD simulations, Mg 2+ ions were placed at different positions as seen in the crystal structures and the movement of the ion was followed over the course of the trajectory. Our analysis suggests a “moving metal mechanism” in which one Mg 2+ ion moves from the B‐ (more buried) to the A‐site during substrate cleavage. The anticipated inversion of the phosphate oxygens occurs during the in‐line cleavage reaction. Experimental results, which show competition between Ca 2+ and Mg 2+ for catalyzing the reaction, and high concentrations of Mg 2+ are inihibitory, indicate that both sites cannot be simultaneously occupied for maximal activity. Proteins 2007. © 2007 Wiley‐Liss, Inc.