Mutations in the proteolytic domain of Escherichia coli protease Lon impair the ATPase activity of the enzyme

Natalie N. Starkova, Ekaterina P. Koroleva, Lev D. Rumsh, Lev M. Ginodman, Tatyana V. Rotanova

Research output: Contribution to journalArticle

45 Scopus citations

Abstract

Conserved residues of the proteolytic domain of Escherichia coli protease Lon, putative members of the classic catalytic triad (H665, H667, D676, and D743) mere identified by comparison of amino acid sequences of Lon proteases. Mutant enzymes containing substitutions D676N, D743N, H665Y, and H667Y were obtained by site-directed mutagenesis. The mutant D743N retained the adenosine triphosphate (ATP)-dependent proteolytic activity, thereby indicating that D743 does not belong to the catalytic site. Simultaneously, the mutants D676N, H665Y, add H667Y lost the capacity for hydrolysis of protein substrates. The ATPase activity of these three mutants was decreased by more than an order of magnitude, which suggests a close spatial location of the ATPase and proteolytic active sites and their tight interaction in the process of protein degradation.

Original languageEnglish (US)
Pages (from-to)218-220
Number of pages3
JournalFEBS Letters
Volume422
Issue number2
DOIs
StatePublished - Jan 30 1998
Externally publishedYes

Keywords

  • Active site
  • Adenosine triphosphate-dependent proteolysis
  • Escherichia coli
  • Protease Lon
  • Site-directed mutagenesis

ASJC Scopus subject areas

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Molecular Biology
  • Genetics
  • Cell Biology

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