solubilization and humanization of paraoxonase-1增溶和paraoxonase-1人性化.pdfVIP

Hindawi Publishing Corporation Journal of Lipids Volume 2012, Article ID 610937, 13 pages doi:10.1155/2012/610937 Research Article Solubilization and Humanization of Paraoxonase-1 Mohosin Sarkar,1 Christina Keventzidis Harsch,1 George T. Matic,1 Kathryn Hoffman,1 Joseph R. Norris III,2 Tamara C. Otto,2 David E. Lenz,2 Douglas M. Cerasoli,2 and Thomas J. Magliery1, 3 1 Department of Chemistry, The Ohio State University, Columbus, OH 43210, USA 2 Physiology and Immunology Branch, Research Division, United States Army Medical Research Institute of Chemical Defense, Aberdeen Proving Ground, MD 21010, USA 3 Department of Biochemistry, The Ohio State University, Columbus, OH 43210, USA Correspondence should be addressed to Thomas J. Magliery, magliery.1@ Received 12 January 2012; Revised 26 March 2012; Accepted 26 March 2012 Academic Editor: Alejandro Gugliucci Copyright © 2012 Mohosin Sarkar et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Paraoxonase-1 (PON1) is a serum protein, the activity of which is related to susceptibility to cardiovascular disease and intoxication by organophosphorus (OP) compounds. It may also be involved in innate immunity, and it is a possible lead molecule in the development of a catalytic bioscavenger of OP pesticides and nerve agents. Human PON1 expressed in E. coli is mostly found in the insoluble fraction, which motivated the engineering of soluble variants, such as G2E6, with more than 50 mutations from huPON1. We examined the effect on the solubility, activity, and stability of three sets of mutations designed to solubilize huPON1