The serine acetyltransferase from Escherichia coli. Over-expression, purification and preliminary crystallographic analysis.
An expression vector has been constructed which increases the expression of serine acetyltransferase (SAT) from E. coli to 17% of the soluble cell protein. A novel purification procedure, using dye-affinity chromatography, allows purification of SAT to homogeneity. The enzyme has been crystallised from polyethylene glycol, in the presence of L-cysteine (an inhibitor of SAT). The crystals which diffract to beyond 3.0 A resolution are of the tetragonal spacegroup P4(1)2(1)2 (or P4(3)2(1)2) with cell dimensions a = b = 123 A, c = 79 A. Since ultracentrifugation and gel-filtration experiment indicate that purified SAT is a tetramer, there appears to be one-half tetramer in the asymmetric unit (Vm = 2.55 A3/Da).
Wigley
D B
DB
Department of Biochemistry, University of Leicester, UK.
Derrick
J P
JP
Shaw
W V
WV
D000123:Acetyltransferases / Q000235:genetics / Q000302:isolation & purification* / Q000648:ultrastructure; D002850:Chromatography, Gel; D003001:Cloning, Molecular; D003461:Crystallography; D004926:Escherichia coli / Q000201:enzymology*; D015870:Gene Expression; D015394:Molecular Structure; D012694:Serine; D051050:Serine O-Acetyltransferase
D012694:Serine; D000123:Acetyltransferases; D051050:Serine O-Acetyltransferase