Purification, characterization, and crystallization of an N-hydroxyarylamine O-acetyltransferase from Salmonella typhimurium.
The N-hydroxyarylamine O-acetyltransferase from Salmonella typhimurium has been expressed as a histidine-tagged fusion protein in Escherichia coli and purified to apparent homogeneity using single-step immobilized metal ion chromatography. Sufficient quantities of the purified protein have been obtained to allow its characterization by physical methods including dynamic light scattering and electrospray mass spectrometry. The substrate specificity and temperature sensitivity of the enzymatic activity have also been assessed. The enzyme has been crystallized from sodium, potassium tartrate and X-ray diffraction data have been obtained to allow the identification of an orthorhombic unit cell, point group P21212, with dimensions a = 137 A, b = 223 A, and c = 105 A. These crystals will provide a route to a crystallographic determination of the structure of the protein.
Sinclair
J C
JC
Department of Pharmacology, Oxford University, Mansfield Road, Oxford, OX1 3QT, England.
Delgoda
R
R
Noble
M E
ME
Jarmin
S
S
Goh
N K
NK
Sim
E
E
D000123:Acetyltransferases; D000217:Acyltransferases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D000814:Aniline Compounds / Q000378:metabolism; D001483:Base Sequence; D017931:DNA Primers / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics*; D007526:Isoelectric Point; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012486:Salmonella typhimurium / Q000201:enzymology* / Q000235:genetics; D013379:Substrate Specificity; D013696:Temperature; D013917:Thrombin / Q000378:metabolism; D014961:X-Ray Diffraction
D000814:Aniline Compounds; D017931:DNA Primers; D011993:Recombinant Fusion Proteins; C013067:4-iodoaniline; D000217:Acyltransferases; D000123:Acetyltransferases; C019736:N-hydroxyarylamine O-acetyltransferase; D013917:Thrombin