Crystallization and preliminary crystallographic analysis of human muscle phosphofructokinase, the main regulator of glycolysis

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...

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Published in:Acta crystallographica. Section F, Structural biology communications Structural biology communications, 2014-05, Vol.70 (5), p.578-582
Main Authors: Kloos, Marco, Brüser, Antje, Kirchberger, Jürgen, Schöneberg, Torsten, Sträter, Norbert
Format: Article
Language:English
Subjects:
allosteric regulation
Amino Acid Sequence
Crystallization
Crystallography
Glycolysis - physiology
human muscle phosphofructokinase
Humans
Molecular Sequence Data
Muscle, Skeletal - enzymology
Phosphofructokinase-1, Muscle Type - chemistry
Phosphofructokinase-1, Muscle Type - physiology
Protein Structure, Secondary
Protein Structure, Tertiary
Structural Communications
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Summary: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.
ISSN:2053-230X
2053-230X
DOI:10.1107/S2053230X14008723