Structural analysis and functional study of phosphofructokinase B (PfkB) from Mycobacterium marinum

Biochem Biophys Res Commun. 2021 Nov 19:579:129-135. doi: 10.1016/j.bbrc.2021.09.051. Epub 2021 Sep 23.

Abstract

Phosphofructokinase B (PfkB) belongs to the ribokinase family, which uses the phosphorylated sugar as substrate, and catalyzes fructose-6-phosphate into fructose-1,6-diphosphate. However, the structural basis of Mycobacterium marinum PfkB is not clear. Here, we found that the PfkB protein was monomeric in solution, which was different from most enzymes in this family. The crystal structure of PfkB protein from M. marinum was solved at a resolution of 2.21 Å. The PfkB structure consists of two domains, a major three-layered α/β/α sandwich-like domain characteristic of the ribokinase-like superfamily, and a second domain composed of four-stranded β sheets. Structural comparison analysis suggested that residues G236, A237, G238, and D239 could be critical for ATP catalysis and substrate binding of PfkB. Our current work provides new insights into understanding the mechanism of the glycolysis in M. marinum.

Keywords: Crystal structure; Mycobacterium marinum; PfkB; Phosphofructokinase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Catalysis
  • Chromatography, Gel
  • Crystallography, X-Ray
  • Escherichia coli
  • Fructosephosphates / chemistry
  • Glycolysis
  • Hydrogen-Ion Concentration
  • Molecular Conformation
  • Molecular Docking Simulation
  • Mycobacterium marinum / enzymology*
  • Phosphofructokinase-2 / metabolism*
  • Phosphotransferases (Alcohol Group Acceptor) / chemistry
  • Protein Conformation
  • Protein Folding
  • Protein Structure, Secondary
  • Scattering, Radiation
  • Temperature

Substances

  • Fructosephosphates
  • fructose-6-phosphate
  • Phosphotransferases (Alcohol Group Acceptor)
  • Phosphofructokinase-2
  • ribokinase