Accession Number : AD1019022


Title :   Expression of Membrane-Bound Human AminopeptidaseP as a Soluble Enzyme and an Investigation into Its Efficacy Towards Offering Protection Against the Toxicity of Chemical Warfare Nerve Agents


Descriptive Note : Technical Report,01 Jan 2013,31 Mar 2015


Corporate Author : ARMY MEDICAL RESEARCH INST OF CHEMICAL DEFENSE ABERDEEN PROVING GROUND MD ABERDEEN PROVING GROUND United States


Personal Author(s) : Sabnekar,Praveena ; Mata,David G ; Rezk,Peter ; Chilukuri,Nageswararao


Full Text : https://apps.dtic.mil/dtic/tr/fulltext/u2/1019022.pdf


Report Date : 01 Sep 2016


Pagination or Media Count : 18


Abstract : In humans there are two forms of aminopeptidaseP (APP), a cytosolic and a membrane-bound (hmAPP). In this study, we produced an adenovirus containing the hmAPP gene without the membrane binding domain and containing a C-terminus 6 histidine tag (AdshmAPP);this virus was used for expression of soluble hmAPP in HEK293A cells in vitro and in mice in vivo. The enzyme fromHEK293A cells was purified and investigated for its ability to hydrolyze GD, GB, GF, GA, and VX. While unable to hydrolyze VX, shmAPP was capable of hydrolyzing each G agent, hydrolyzing GD more efficiently than any other G-type nerve agent with a catalytic efficiency of (5 1) 106 M-1 min-1. However, the stereo chemical preference of shmAPP favors the hydrolysis of the non-toxic P (+)isomer(s) of each agent. We then evaluated whether mice containing elevated blood levels of shmAPP would be protected from the lethality of GD, GF and GA. Mice were infected with Ad-shmAPP and 5 days later were challenged subcutaneously with 1 x LD50 doses of GA,GF and GD. The survival rates of Ad-shmAPP-treated mice were not significantly different from those of mice treated with a control virus. These results suggest that the overexpression of wild-type shmAPP in mice failed to afford protection against nerve agent toxicity.


Descriptors :   chemical warfare agents , gene expression , medical countermeasures , toxicity , nerve agents , enzymes


Subject Categories : Chemical, Biological and Radiological Warfare


Distribution Statement : APPROVED FOR PUBLIC RELEASE