title: Archaeal ADP-dependent phosphofructokinase: expression, purification, crystallization and preliminary crystallographic analysis.
abstract: Thermococcus litoralis uses a modified Embden-Meyerhof pathway. Its phosphofructokinase (TLPFK) catalyses the phosphorylation of fructose-6-phosphate by ADP, but not by ATP, in the presence of Mg(2+), yielding fructose-1,6-bisphosphate. The gene encoding TLPFK was cloned and overexpressed in Escherichia coli. Recombinant TLPFK consists of a dimer with a 52 kDa subunit. The native crystals belong to space group P4(1)2(1)2, with unit-cell parameters a = b = 85.41, c = 163.93 A, and diffract to beyond 2.6 A resolution. The TLPFK structure was preliminarily analyzed by means of multiple isomorphous replacement with four heavy-atom derivatives.
authors.affiliation: Department of Biotechnology, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
authors.forename: Jong-Jin
authors.initials: JJ
authors.lastname: Jeong
authors.forename: Shinya
authors.initials: S
authors.lastname: Fushinobu
authors.forename: Sohei
authors.initials: S
authors.lastname: Ito
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Shoun
authors.forename: Takayoshi
authors.initials: T
authors.lastname: Wakagi
mesh_terms: D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D004269:DNA, Bacterial / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics; D008969:Molecular Sequence Data; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000737:chemistry; D017422:Sequence Analysis, DNA; D019712:Thermococcus / Q000201:enzymology* / Q000235:genetics
chemical_list: D004269:DNA, Bacterial; D011994:Recombinant Proteins; D017853:Phosphotransferases (Alcohol Group Acceptor); C442129:ADP D-fructose-6-phosphate 1-phosphotransferase