title: Lysine carboxylation in proteins: OXA-10 beta-lactamase.
abstract: An increasing number of proteins are being shown to have an N(zeta)-carboxylated lysine in their structures, a posttranslational modification of proteins that proceeds without the intervention of a specific enzyme. The role of the carboxylated lysine in these proteins is typically structural (hydrogen bonding or metal coordination). However, carboxylated lysines in the active sites of OXA-10 and OXA-1 beta-lactamases and the sensor domain of BlaR signal-transducer protein serve in proton transfer events required for the functions of these proteins. These examples demonstrate the utility of this unusual amino acid in acid-base chemistry, in expansion of function beyond those of the 20 standard amino acids. In this study, the ONIOM quantum-mechanical/molecular-mechanical (QM/MM) method is used to study the carboxylation of lysine in the OXA-10 beta-lactamase. Lys-70 and the active site of the OXA-10 beta-lactamase were treated with B3LYP/6-31G(d,p) density functional calculations and the remainder of the enzyme with the AMBER molecular mechanics force field. The barriers for unassisted carboxylation of neutral lysine by carbon dioxide or bicarbonate are high. However, when the reaction with CO2 is catalyzed by a molecule of water in the active site, it is exothermic by about 13 kcal/mol, with a barrier of approximately 14 kcal/mol. The calculations show that the carboxylation and decarboxylation of Lys-70 are likely to be accompanied by deprotonation and protonation of the carbamate, respectively. The analysis may also be relevant for other proteins with carboxylated lysines, a feature that may be more common in nature than previously appreciated.
authors.affiliation: Department of Chemistry and Institute for Scientific Computing, Wayne State University, Detroit, Michigan 48202, USA.
authors.forename: Jie
authors.initials: J
authors.lastname: Li
authors.forename: Jason B
authors.initials: JB
authors.lastname: Cross
authors.forename: Thom
authors.initials: T
authors.lastname: Vreven
authors.forename: Samy O
authors.initials: SO
authors.lastname: Meroueh
authors.forename: Shahriar
authors.initials: S
authors.lastname: Mobashery
authors.forename: H Bernhard
authors.initials: HB
authors.lastname: Schlegel
mesh_terms: D019295:Computational Biology; D008239:Lysine / Q000737:chemistry* / Q000378:metabolism; D008744:Methylamines / Q000737:chemistry / Q000378:metabolism; D015394:Molecular Structure; D011499:Protein Processing, Post-Translational; D001618:beta-Lactamases / Q000737:chemistry* / Q000378:metabolism
chemical_list: D008744:Methylamines; C027451:methylamine; C425107:beta-lactamase OXA-10; D001618:beta-Lactamases; D008239:Lysine