Use of chemical modification in the crystallization of isocitrate lyase from Escherichia coli.
Two different crystal forms of isocitrate lyase (ICL) from Escherichia coli have been grown following the chemical modification of the enzyme by either 3-bromopyruvate or ethyl mercuri thiosalicylate (EMTS), contrasting strongly with difficulties in obtaining ordered crystals of the native enzyme. Both crystal forms are obtained using the hanging drop method of vapour diffusion with ammonium sulphate as the precipitant. The crystals diffract well and X-ray photographs of the crystals have established that they are in space groups C222(1) and P3(1) (or its enantiomorph P3(2), respectively. Considerations of the values of Vm and measurements on the crystal density indicate that the asymmetric unit of both crystals contains four subunits.
Abeysinghe
S I
SI
Krebs Institute for Biomolecular Research, Department of Molecular Biology and Biotechnology, University of Sheffield, U.K.
Baker
P J
PJ
Rice
D W
DW
Rodgers
H F
HF
Stillman
T J
TJ
Ko
Y H
YH
McFadden
B A
BA
Nimmo
H G
HG
D003460:Crystallization; D004926:Escherichia coli / Q000201:enzymology*; D007522:Isocitrate Lyase / Q000737:chemistry* / Q000302:isolation & purification; D046911:Macromolecular Substances; D011487:Protein Conformation; D011773:Pyruvates / Q000494:pharmacology*; D013849:Thimerosal / Q000494:pharmacology*; D014961:X-Ray Diffraction
D046911:Macromolecular Substances; D011773:Pyruvates; D013849:Thimerosal; C017092:bromopyruvate; D007522:Isocitrate Lyase