Purification, characterization and crystallization of thermostable anthranilate phosphoribosyltransferase from Sulfolobus solfataricus.
Anthranilate phosphoribosyltransferase (TrpD; EC 2.4.2.18) from the hyperthermophilic archaeon Sulfolobus solfataricus (ssTrpD) was expressed in Escherichia coli, purified and crystallized. Analytical gel permeation chromatography revealed a homodimeric composition of the enzyme. The steady-state kinetic characteristics suggest tight binding of the substrate anthranilic acid and efficient catalysis at the physiological growth temperature of S. solfataricus. Crystals of ssTrpD diffract to better than 2.6 A resolution and preliminary X-ray characterization was carried out. The crystals are suitable for structure determination.
Ivens
A
A
Institut für Biochemie der Universität zu Köln, Germany; EMBL-Hamburg Outstation, Hamburg, Germany. andreas.ivens@uni-koeln.de
Mayans
O
O
Szadkowski
H
H
Wilmanns
M
M
Kirschner
K
K
D000877:Anthranilate Phosphoribosyltransferase / Q000737:chemistry* / Q000302:isolation & purification*; D002384:Catalysis; D002850:Chromatography, Gel; D003461:Crystallography; D018360:Crystallography, X-Ray; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism; D007700:Kinetics; D008715:Methionine / Q000737:chemistry; D008956:Models, Chemical; D011485:Protein Binding; D016944:Sulfolobus / Q000201:enzymology*; D013696:Temperature; D013997:Time Factors; D062367:ortho-Aminobenzoates / Q000737:chemistry
D062367:ortho-Aminobenzoates; C031385:anthranilic acid; D008715:Methionine; D000877:Anthranilate Phosphoribosyltransferase