title: Crystallization and preliminary crystallographic analysis of human muscle phosphofructokinase, the main regulator of glycolysis.
abstract: Whereas the three-dimensional structure and the structural basis of the allosteric regulation of prokaryotic 6-phosphofructokinases (Pfks) have been studied in great detail, knowledge of the molecular basis of the allosteric behaviour of the far more complex mammalian Pfks is still very limited. The human muscle isozyme was expressed heterologously in yeast cells and purified using a five-step purification protocol. Protein crystals suitable for diffraction experiments were obtained by the vapour-diffusion method. The crystals belonged to space group P6222 and diffracted to 6.0 Å resolution. The 3.2 Å resolution structure of rabbit muscle Pfk (rmPfk) was placed into the asymmetric unit and optimized by rigid-body and group B-factor refinement. Interestingly, the tetrameric enzyme dissociated into a dimer, similar to the situation observed in the structure of rmPfk.
authors.affiliation: Institute of Bioanalytical Chemistry, Center for Biotechnology and Biomedicine, University of Leipzig, Deutscher Platz 5, 04103 Leipzig, Germany.
authors.forename: Marco
authors.initials: M
authors.lastname: Kloos
authors.affiliation: Institute of Biochemistry, Medical Faculty, University of Leipzig, Johannisallee 30, 04103 Leipzig, Germany.
authors.forename: Antje
authors.initials: A
authors.lastname: Brüser
authors.affiliation: Institute of Biochemistry, Medical Faculty, University of Leipzig, Johannisallee 30, 04103 Leipzig, Germany.
authors.forename: Jürgen
authors.initials: J
authors.lastname: Kirchberger
authors.affiliation: Institute of Biochemistry, Medical Faculty, University of Leipzig, Johannisallee 30, 04103 Leipzig, Germany.
authors.forename: Torsten
authors.initials: T
authors.lastname: Schöneberg
authors.affiliation: Institute of Bioanalytical Chemistry, Center for Biotechnology and Biomedicine, University of Leipzig, Deutscher Platz 5, 04103 Leipzig, Germany.
authors.forename: Norbert
authors.initials: N
authors.lastname: Sträter
mesh_terms: D000595:Amino Acid Sequence; D003460:Crystallization; D003461:Crystallography; D006019:Glycolysis / Q000502:physiology*; D006801:Humans; D008969:Molecular Sequence Data; D018482:Muscle, Skeletal / Q000201:enzymology*; D025203:Phosphofructokinase-1, Muscle Type / Q000737:chemistry* / Q000502:physiology*; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary
chemical_list: D025203:Phosphofructokinase-1, Muscle Type
keywords: allosteric regulation; human muscle phosphofructokinase