Crystallization and preliminary X-ray crystallographic studies on recombinant rat choline acetyltransferase.
Choline acetyltransferase (ChAT) catalyzes the biosynthesis of the neurotransmitter acetylcholine from acetyl-CoA and choline in cholinergic neurons. Rat ChAT (rChAT) was overexpressed in Escherichia coli, purified by affinity chromatography and crystallized. Diffraction data were collected from a single crystal under cryoconditions at the F1 beamline at the Cornell High Energy Synchrotron Source, with a maximal useful diffraction pattern to 1.55 A resolution. The crystals were shown to belong to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 138.97, b = 77.67, c = 59.67 A and a scaling R(sym) of 0.054 for 72 446 unique reflections. Packing considerations indicate there to be one molecule per asymmetric unit. It is expected that in the near future the structure of rChAT will be obtained using molecular-replacement methods. Elucidation of the structure of rChAT will aid in the development of therapeutic agents for Alzheimer's disease.
Lian
Wei
W
Department of Medicinal Chemistry, McKnight Brain Institute and University of Florida, Gainesville, Florida 32610, USA.
Gu
Yunrong
Y
Pedersen
Brenda
B
Kukar
Thomas
T
Govindasamy
Lakshmanan
L
Agbandje-McKenna
Mavis
M
Jin
Shouguang
S
McKenna
Robert
R
Wu
Donghai
D
D000818:Animals; D002795:Choline O-Acetyltransferase / Q000737:chemistry*; D003460:Crystallization; D018360:Crystallography, X-Ray / Q000379:methods*; D004926:Escherichia coli / Q000378:metabolism; D009474:Neurons / Q000378:metabolism; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry*; D017356:Synchrotrons
D011994:Recombinant Proteins; D002795:Choline O-Acetyltransferase