Structural and biochemical characterization of inorganic pyrophosphatase from Homo sapiens

Biochem Biophys Res Commun. 2020 Dec 17;533(4):1115-1121. doi: 10.1016/j.bbrc.2020.09.139. Epub 2020 Oct 6.

Abstract

Inorganic pyrophosphatase (PPase) plays an essential role in energy conservation and provides energy for many biosynthetic pathways. Here, we present two three-dimensional structures of PPase from Homo sapiens (Hu-PPase) at 2.38 Å and 3.40 Å in different crystallization conditions. One of the Hu-PPase structures complex of two magnesium metal ions was determined to be a monomer (Hu-PPase-mono) here, while the other one to be a dimer-dimer (Hu-PPase-dd). In each asymmetric unit of Hu-PPase-mono, there are four α-helices and ten β-strands and folds as a barrel structure, and the active site contains two magnesium ions. Like PPases from many species, we found that Hu-PPase was able to undergo self-assembly. To our surprise, disruption of the self-assembly of Hu-PPase did not influence its enzymatic activity or the ability to promote cell growth. Our work uncovered that different structure forms of Hu-PPase and found that the pyrophosphatase activity of Hu-PPase is independent of its self-assembly.

Keywords: Cell proliferation; Crystal structure; Hu-PPase; Pyrophosphatase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Binding Sites
  • Catalysis
  • Cell Proliferation / genetics*
  • Crystallography, X-Ray
  • Dimerization
  • HEK293 Cells
  • HeLa Cells
  • Humans
  • Inorganic Pyrophosphatase / chemistry*
  • Inorganic Pyrophosphatase / genetics
  • Inorganic Pyrophosphatase / metabolism*
  • Magnesium / chemistry*
  • Models, Molecular
  • Protein Conformation
  • Protein Conformation, alpha-Helical
  • Protein Conformation, beta-Strand
  • Recombinant Proteins

Substances

  • Recombinant Proteins
  • Inorganic Pyrophosphatase
  • Magnesium