Design and Production of an Organophosphorus-Bioscavenger via Protein Engineering of the Human Acetylcholinesterase
Abstract
The design of novel biocatalysts with potential pharmacological use against organophosphate (OP) poisoning, within the mold of recombinant human acetyicholinesterase (rHuAChE), was carried out by two complementing research approaches: a.) Kinetic studies combined with mass spectrometric analysis of rHuAChE and selected mutants thereof, with a variety of OPs for better understanding of the chemical environment for the OPs in the active center of AChE. Specifically, here we describe the effects of multiple replacements of aromatic residues at the active center on reactivity toward OP agents, as well as report on the resolution of some of the less understood reaction pathways of phosphylation, and "aging" involving phosphoroamidates (P-N agents) such as tabun and butyl-tabun. b.) Analyses of post-translation modifications responsible for the limited residence time of rHuAChE: Extensive structural analysis of the N-glycans of recombinant bovine and human AChE and native bovine AChE, identified specific termini that account for the observed differential behavior of the recombinant and native enzymes.
Document Details
- Document Type
- Technical Report
- Publication Date
- Sep 01, 2002
- Accession Number
- ADA416886
Entities
People
- Avigdor Shafferman
Organizations
- Israel Institute for Biological Research