Functional analysis of propeptide as an intramolecular chaperone for in vivo folding of subtilisin nattokinase

FEBS Lett. 2010 Dec 1;584(23):4789-96. doi: 10.1016/j.febslet.2010.11.011. Epub 2010 Nov 11.

Abstract

Here, we show that during in vivo folding of the precursor, the propeptide of subtilisin nattokinase functions as an intramolecular chaperone (IMC) that organises the in vivo folding of the subtilisin domain. Two residues belonging to β-strands formed by conserved regions of the IMC are crucial for the folding of the subtilisin domain through direct interactions. An identical protease can fold into different conformations in vivo due to the action of a mutated IMC, resulting in different kinetic parameters. Some interfacial changes involving conserved regions, even those induced by the subtilisin domain, blocked subtilisin folding and altered its conformation. Insight into the interaction between the subtilisin and IMC domains is provided by a three-dimensional structural model.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Bacillus subtilis / enzymology
  • Conserved Sequence
  • Kinetics
  • Molecular Chaperones / chemistry
  • Molecular Chaperones / genetics
  • Molecular Chaperones / metabolism*
  • Molecular Dynamics Simulation
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • Mutation
  • Protein Folding*
  • Protein Precursors / chemistry
  • Protein Precursors / genetics
  • Protein Precursors / metabolism*
  • Protein Refolding
  • Protein Structure, Secondary
  • Protein Structure, Tertiary
  • Sequence Alignment
  • Sequence Homology, Amino Acid
  • Subtilisins / chemistry*
  • Subtilisins / genetics
  • Subtilisins / metabolism*

Substances

  • Molecular Chaperones
  • Protein Precursors
  • Subtilisins
  • nattokinase