1. Mn2+ modulates the kinetic properties of an archaeal member of the PLL family
- Author
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Spartaco Di Gennaro, Elena Porzio, Giuseppe Manco, and Achille Palma
- Subjects
Sulfolobus acidocaldarius ,STRUCTURAL BASIS ,LACTONASE ,Stereochemistry ,DIRECTED EVOLUTION ,Archaeal Proteins ,Gene Expression ,NERVE AGENTS ,Toxicology ,Mass Spectrometry ,Paraoxon ,Genes, Archaeal ,Substrate Specificity ,Metal ,THERMOSTABLE PHOSPHOTRIESTERASE ,Apoenzymes ,AMIDOHYDROLASE SUPERFAMILY ,Enzyme Stability ,Lactonase ,medicine ,chemistry.chemical_classification ,Manganese ,DEINOCOCCUS-RADIODURANS ,biology ,Amidohydrolase ,ACTIVE-SITE ,PSEUDOMONAS-DIMINUTA ,Temperature ,Substrate (chemistry) ,General Medicine ,Hydrogen-Ion Concentration ,Recombinant Proteins ,Kinetics ,Enzyme ,Phosphoric Triester Hydrolases ,Biochemistry ,chemistry ,visual_art ,biology.protein ,visual_art.visual_art_medium ,SULFOLOBUS-SOLFATARICUS ,Holoenzymes ,Carboxylic Ester Hydrolases ,Function (biology) ,medicine.drug ,Cadmium - Abstract
Recently we reported on the characterization of an archaeal member of the amidohydrolase superfamily, namely Sulfolobus acidocaldarius lactonase, showing low but significant and extremely thermostable paraoxonase activity. This enzyme, that we have named SacPox, is a member of the new described family of phosphotriesterase-like lactonases (PLLs). In this family the binuclear metal centre, which is involved in the catalytic machinery, has been poorly studied up to now. In this work we describe the expression of the protein in presence of different metals showing Mn2+ to support the higher activity. The enzyme has been over-expressed, purified and characterized as a Mn2+-containing enzyme by inductive plasma coupled mass spectrometry (ICP-MS), showing also surprising kinetic differences in comparison with the cadmium-containing enzyme. The Mn2+ containing enzyme was about 30-fold more efficient with paraoxon as substrate and more stable than the Cd2+ counterpart, even though the Mn2+ affinity for the binuclear metal centre is apparently lower. These results increase our knowledge of the biochemical characteristics of SacPox mainly with regard to the metal-ions modulation of function. (C) 2012 Elsevier Ireland Ltd. All rights reserved.
- Published
- 2012
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