Crystallization and structure solution at 4 A resolution of the recombinant synthase domain of N-(5'-phosphoribosyl)anthranilate isomerase:indole-3-glycerol-phosphate synthase from Escherichia coli complexed to a substrate analogue.
The recombinant synthase domain of the bifunctional enzyme N-(5'-phosphoribosyl)anthranilate isomerase:indole-3-glycerol-phosphate synthase from Escherichia coli has been crystallized, and the structure has been solved at 4 A resolution. Two closely related crystal forms grown from ammonium sulphate diffract to 2 A resolution. One form (space group R32, a = 163 A, alpha = 29.5 degrees) contains the unliganded synthase domain; the second crystal form (space group P6(3)22, a = 144 A, c = 158 A) is co-crystallized with the substrate analogue N-(5'-phosphoribit-1-yl)anthranilate. The structure of the synthase-inhibitor complex has been solved by the molecular replacement method. This achievement represents the first successful use of a (beta alpha)8-barrel monomer as a trial model. The recombinant synthase domain associates as a trimer in the crystal, the molecules being related by a pseudo-crystallographic triad. The interface contacts between the three domains are mediated by those residues that are also involved in the domain interface of the bifunctional enzyme. This system provides a model for an interface which is used in both intermolecular and intramolecular domain contacts.
Wilmanns
M
M
Department of Structural Biology, University of Basel, Switzerland.
Schlagenhauf
E
E
Fol
B
B
Jansonius
J N
JN
D019747:Aldose-Ketose Isomerases; D002238:Carbohydrate Epimerases / Q000378:metabolism; D002262:Carboxy-Lyases / Q000378:metabolism; D003460:Crystallization; D004228:Dithionitrobenzoic Acid; D004926:Escherichia coli / Q000201:enzymology*; D007209:Indole-3-Glycerol-Phosphate Synthase / Q000378:metabolism; D046911:Macromolecular Substances; D015394:Molecular Structure; D011994:Recombinant Proteins; D012267:Ribosemonophosphates / Q000378:metabolism; D014961:X-Ray Diffraction; D062367:ortho-Aminobenzoates / Q000378:metabolism
D046911:Macromolecular Substances; D011994:Recombinant Proteins; D012267:Ribosemonophosphates; D062367:ortho-Aminobenzoates; C063777:N-(5'-phosphoribosyl)anthranilate; D004228:Dithionitrobenzoic Acid; D002262:Carboxy-Lyases; D007209:Indole-3-Glycerol-Phosphate Synthase; D002238:Carbohydrate Epimerases; D019747:Aldose-Ketose Isomerases; C029865:phosphoribosylanthranilate isomerase