The Crystallization of Acetylcholinesterase (AChE) from Torpedo Electric Organ
Abstract
The objective of this project is the crystallization of the enzyme acetylcholinesterase (AChE), with the long-term objective of determining its three-dimensional structure and, thereby, the detailed topography of its active site. Torpedo electric organ was selected since it is a rich source of AChE and possesses an amino acid sequence very similar to that of mammalian AChE. A dimeric form of this enzyme was purified by a procedure which involved selective solubilization with a phosphatidylinositol-specific phospholipase C of bacterial origin, followed by affinity chromatography employing a Sepharose conjugate of a suitable quaternary affinity ligand. A highly purified AChE preparation was obtained in amounts which permitted a systematic attempt to crystallize the enzyme. In order to obtain a crystal form of the ACHE preparation suitable for high-resolution X-ray studies, we examined hundreds of different crystallization conditions. As a result we were able to obtain three (3) different crystal forms which diffract to better than 3 A resolution.... RAV, Acetylcholinesterase, X- Ray Crystallography, Torpedo californica, Crystallization, Three-dimensional structure, Enzymes, Acetylcholine.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jul 18, 1991
- Accession Number
- ADA268487
Entities
People
- I. Silman
- J. L. Sussman
Organizations
- Weizmann Institute of Science