Abstract
A knowledge of the 3D structure of human acetylcholinesterase (AChE) is of importance for the development of anti-Alzheimer drugs, for the general understanding of organophosphate toxicity and for the design of safer and more specific insecticides. A mutant of recombinant human acetylcholinesterase (rhAChE) was constructed in which the C-terminus had been truncated to give rise to a homogeneous monomeric form. The monomeric rhAChE was expressed in HEK 293 cells and purified by affinity chromatog-raphy. The purified enzyme was co-crystallized from ammonium sulfate at pH 7.2 and 19°C as a stoichiometric complex with the mamba venom polypeptide toxin, fasciculin-II (FAS-II). X-ray data were collected at the BNL-NSLS X12-C source from a single cryogenically cooled crystal. The overall completeness of the 30A-2.7Å data is 99.0% and the R-merge is 8.1%. The model was refined starting from the mouse AChE-FAS-II structure to Rfree of 29% and R of 21.9% at 2.7Å resolution. The overall fold is similar to that of Torpedo californica and mouse complexes with FAS-II. A mutant E202(199)Q was prepared and crystallized in a similar fashion. Although significantly different from the native enzyme in activity, the E202(199)Q is almost identical to the native in its 3D structure.
Keywords
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsAuthor information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1998 Springer Science+Business Media New York
About this chapter
Cite this chapter
Kryger, G. et al. (1998). 3D Structure of a Complex of Human Acetylcholinesterase with Fasciculin-II at 2.7 Å Resolution. In: Doctor, B.P., Taylor, P., Quinn, D.M., Rotundo, R.L., Gentry, M.K. (eds) Structure and Function of Cholinesterases and Related Proteins. Springer, Boston, MA. https://doi.org/10.1007/978-1-4899-1540-5_102
Download citation
DOI: https://doi.org/10.1007/978-1-4899-1540-5_102
Publisher Name: Springer, Boston, MA
Print ISBN: 978-1-4899-1542-9
Online ISBN: 978-1-4899-1540-5
eBook Packages: Springer Book Archive