Production and Enzyme Engineering of Human Acetyicholinesterase and Its Mutant Derivatives.
Abstract
The research is directed towards design of pharmacologically useful bio-scavengers against organophosphate poisoning, by: (a) structure-reactivity studies of human acetylcholinesterase (HuAChE) and its mutant derivatives with various covalent and noncovalent ligands including CW agents; and (b) studies on molecular surface properties of HuAChE derivatives affecting biosynthesis, stability and clearance. Combination of recombinant DNA technology, kinetic studies , molecular modeling, x-ray crystallography and electro-spray MS methods are employed to identify the residues involved in direct interactions with phosphates and phosphonates and in the subsequent reactivation or aging processes. Our findings allow to define the structural determinants in the active center facilitating the bio-scavenging and provide clear directions for design of optimal bioscavengers which will combine advantageous properties of phosphylation, reactivation and aging via targeting mutations at specific residues. In the past we have demonstrated that a control over the number of vacant sialic acid attachment sites may improve the enzyme residence time in the bloodstream. Here we report that indeed by engineering cells expressing high levels of both recombinant HuAChE and recombinant 2,6-sialyltransferase we were able to extend significantly the residence time of the HuAChE bioscavenger.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jun 01, 1998
- Accession Number
- ADA352171
Entities
People
- Avigdor Shafferman
Organizations
- Israel Institute for Biological Research