1WEI
Catalytic Domain Of Muty From Escherichia Coli K20A Mutant Complexed To Adenine
Summary for 1WEI
| Entry DOI | 10.2210/pdb1wei/pdb |
| Related | 1WEF 1WEG |
| Descriptor | A/G-specific adenine glycosylase, ADENINE, 1,2-ETHANEDIOL, ... (5 entities in total) |
| Functional Keywords | hydrolase |
| Biological source | Escherichia coli |
| Total number of polymer chains | 1 |
| Total formula weight | 25663.86 |
| Authors | Hitomi, K.,Arvai, A.S.,Tainer, J.A. (deposition date: 2004-05-25, release date: 2004-09-21, Last modification date: 2024-05-29) |
| Primary citation | Manuel, R.C.,Hitomi, K.,Arvai, A.S.,House, P.G.,Kurtz, A.J.,Dodson, M.L.,McCullough, A.K.,Tainer, J.A.,Lloyd, R.S. Reaction intermediates in the catalytic mechanism of Escherichia coli MutY DNA glycosylase J.Biol.Chem., 279:46930-46939, 2004 Cited by PubMed Abstract: The Escherichia coli adenine DNA glycosylase, MutY, plays an important role in the maintenance of genomic stability by catalyzing the removal of adenine opposite 8-oxo-7,8-dihydroguanine or guanine in duplex DNA. Although the x-ray crystal structure of the catalytic domain of MutY revealed a mechanism for catalysis of the glycosyl bond, it appeared that several opportunistically positioned lysine side chains could participate in a secondary beta-elimination reaction. In this investigation, it is established via site-directed mutagenesis and the determination of a 1.35-A structure of MutY in complex with adenine that the abasic site (apurinic/apyrimidinic) lyase activity is alternatively regulated by two lysines, Lys142 and Lys20. Analyses of the crystallographic structure also suggest a role for Glu161 in the apurinic/apyrimidinic lyase chemistry. The beta-elimination reaction is structurally and chemically uncoupled from the initial glycosyl bond scission, indicating that this reaction occurs as a consequence of active site plasticity and slow dissociation of the product complex. MutY with either the K142A or K20A mutation still catalyzes beta and beta-delta elimination reactions, and both mutants can be trapped as covalent enzyme-DNA intermediates by chemical reduction. The trapping was observed to occur both pre- and post-phosphodiester bond scission, establishing that both of these intermediates have significant half-lives. Thus, the final spectrum of DNA products generated reflects the outcome of a delicate balance of closely related equilibrium constants. PubMed: 15326180DOI: 10.1074/jbc.M403944200 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (1.45 Å) |
Structure validation
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