title: Hydrolysis potential of recombinant human skin and kidney prolidase against diisopropylfluorophosphate and sarin by in vitro analysis.
abstract: Human prolidase (PROL), which has structural homology to bacterial organophosphate acid anhydrolase that hydrolyze organophosphates and nerve agents has been proposed recently as a potential catalytic bioscavenger. To develop PROL as a catalytic bioscavenger, we evaluated the in vitro hydrolysis efficiency of purified recombinant human PROL against organophosphates and nerve agents. Human liver PROL was purified by chromatographic procedures, whereas recombinant human skin and kidney PROL was expressed in Trichoplusia ni larvae, affinity purified and analyzed by gel electrophoresis. The catalytic efficiency of PROL against diisopropylfluorophosphate (DFP) and nerve agents was evaluated by acetylcholinesterase back-titration assay. Partially purified human liver PROL hydrolyzed DFP and various nerve agents, which was abolished by specific PROL inhibitor showing the specificity of hydrolysis. Both the recombinant human skin and kidney PROL expressed in T. ni larvae showed ∼99% purity and efficiently hydrolyzed DFP and sarin. In contrast to human liver PROL, both skin and kidney PROL showed significantly low hydrolyzing potential against nerve agents soman, tabun and VX. In conclusion, compared to human liver PROL, recombinant human skin and kidney PROL hydrolyze only DFP and sarin showing the substrate specificity of PROL from various tissue sources.
authors: Blast-Induced Neurotrauma Branch, Center for Military Psychiatry and Neuroscience, Walter Reed Army Institute of Research, Silver Spring, MD 20910, USA.
authors: Michael
authors: M
authors: Costante
authors: Lionel
authors: L
authors: Biggemann
authors: Yonas
authors: Y
authors: Alamneh
authors: Iswarduth
authors: I
authors: Soojhawon
authors: Radley
authors: R
authors: Short
authors: Savita
authors: S
authors: Nigam
authors: Gregory
authors: G
authors: Garcia
authors: Bhupendra P
authors: BP
authors: Doctor
authors: Manojkumar
authors: M
authors: Valiyaveettil
authors: Madhusoodana P
authors: MP
authors: Nambiar
mesh_terms: D000110:Acetylcholinesterase / Q000737:chemistry; D002619:Chemical Warfare Agents / Q000737:chemistry*; D002800:Cholinesterase Inhibitors / Q000737:chemistry*; D004150:Dipeptidases / Q000737:chemistry*; D006801:Humans; D006868:Hydrolysis; D007531:Isoflurophate / Q000737:chemistry; D007668:Kidney / Q000201:enzymology; D008099:Liver / Q000201:enzymology; D010755:Organophosphates / Q000737:chemistry; D009946:Organothiophosphorus Compounds / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry*; D012524:Sarin / Q000737:chemistry; D012867:Skin / Q000201:enzymology
chemical_list: D000110:Acetylcholinesterase; D002619:Chemical Warfare Agents; D002800:Cholinesterase Inhibitors; D004150:Dipeptidases; D007531:Isoflurophate; D010755:Organophosphates; D009946:Organothiophosphorus Compounds; D011994:Recombinant Proteins; D012524:Sarin; C009680:VX-agent; C022449:proline dipeptidase; C009374:tabun