Lampitella_2024_Molecules_29_

Reference

Title : The Human Paraoxonase 2: An Optimized Procedure for Refolding and Stabilization Facilitates Enzyme Analyses and a Proteomics Approach - Lampitella_2024_Molecules_29_
Author(s) : Lampitella EA , Marone M , Achanta NSK , Porzio E , Trepiccione F , Manco G
Ref : Molecules , 29 : , 2024
Abstract :

The human paraoxonase 2 (PON2) is the oldest member of a small family of arylesterase and lactonase enzymes, representing the first line of defense against bacterial infections and having a major role in ROS-associated diseases such as cancer, cardiovascular diseases, neurodegeneration, and diabetes. Specific Post-Translational Modifications (PTMs) clustering nearby two residues corresponding to pon2 polymorphic sites and their impact on the catalytic activity are not yet fully understood. Thus, the goal of the present study was to develop an improved PON2 purification protocol to obtain a higher amount of protein suitable for in-depth biochemical studies and biotechnological applications. To this end, we also tested several compounds to stabilize the active monomeric form of the enzyme. Storing the enzyme at 4 degreesC with 30 mM Threalose had the best impact on the activity, which was preserved for at least 30 days. The catalytic parameters against the substrate 3-Oxo-dodecanoyl-Homoserine Lactone (3oxoC12-HSL) and the enzyme ability to interfere with the biofilm formation of Pseudomonas aeruginosa (PAO1) were determined, showing that the obtained enzyme is well suited for downstream applications. Finally, we used the purified rPON2 to detect, by the direct molecular fishing (DMF) method, new putative PON2 interactors from soluble extracts of HeLa cells.

PubMedSearch : Lampitella_2024_Molecules_29_
PubMedID: 38893310

Related information

Citations formats

Lampitella EA, Marone M, Achanta NSK, Porzio E, Trepiccione F, Manco G (2024)
The Human Paraoxonase 2: An Optimized Procedure for Refolding and Stabilization Facilitates Enzyme Analyses and a Proteomics Approach
Molecules 29 :

Lampitella EA, Marone M, Achanta NSK, Porzio E, Trepiccione F, Manco G (2024)
Molecules 29 :