Stable monomeric form of an originally dimeric serine proteinase inhibitor, ecotin, was constructed via site directed mutagenesis

G. Pál, László Szilágyi, L. Gráf

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Ecotin, a homodimer protein of E. coli, is a unique member of canonical serine proteinase inhibitors, since it is a potent agent against a variety of serine proteinases having different substrate specificity. Monomers of ecotin are held together mostly by their long C-terminal strands that are arranged as a two-stranded antiparallel β-sheet in the functional dimer. One ecotin dimer can chelate two proteinase molecules, each of them bound to both subunits of ecotin at two different sites, namely the specific primary and the non-specific secondary binding sites. In this study the genes of wild type ecotin and its Met84Arg P1 site mutant were truncated resulting in new forms of ecotin that lack 10 amino acid residues at their C-terminus. These mutants do not dimerize spontaneously, though in combination with trypsin they assemble into the familiar heterotetramer. Our data suggest that this heterotetramer exists even in extremely diluted solutions, and the interaction, which is responsible for the dimerization of ecotin, contributes to the stability of the heterotetrameric complex.

Original languageEnglish
Pages (from-to)165-170
Number of pages6
JournalFEBS Letters
Volume385
Issue number3
DOIs
Publication statusPublished - May 6 1996

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Mutagenesis
Serine Proteinase Inhibitors
Escherichia coli Proteins
Dimerization
Serine Proteases
Substrate Specificity
Site-Directed Mutagenesis
Dimers
Trypsin
Peptide Hydrolases
Binding Sites
Amino Acids
Genes
Monomers
Molecules
Substrates

Keywords

  • ecotin
  • serine proteinase inhibitor
  • site directed mutagenesis

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Stable monomeric form of an originally dimeric serine proteinase inhibitor, ecotin, was constructed via site directed mutagenesis. / Pál, G.; Szilágyi, László; Gráf, L.

In: FEBS Letters, Vol. 385, No. 3, 06.05.1996, p. 165-170.

Research output: Contribution to journalArticle

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