Involvement of glutamic acid residue at position 7 in the formation of the intramolecular disulfide bond of Escherichia coli heat-stable enterotoxin Ip in vivo

Hiroyasu Yamanaka, Tomohiko Nomura, Keinosuke Okamoto

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Escherichia coli heat-stable enterotoxin Ip (STIp) is a small peptide toxin composed of 18 amino acid residues containing three intramolecular disulfide bonds. We found previously that the bonds are formed by the catalysis of DsbA (a oxidoreductase) in periplasm. To interact with DsbA, the STIp in periplasm must have a structure suitable as substrate. However, the amino acid residues contributing to the construction of this structure have not been elucidated. We mutated the codon for the glutamic acid at position 7 of STIp by oligonucleotide site-specific mutagenesis in vivo and analysed the STIp produced from the mutant gene. The intramolecular disulfide bonds were not formed in mutant STIp(Glu-7→Ala), but were formed in mutant STIp(Glu-7→Asp). Furthermore, we found that replacing the asparagine residue at position 11 and the proline residue at position 12 did not affect the disulfide bond formation of STIp. The results indicate that a negative charge at position 7 in the sequence of STIp is necessary for STIp to interact with DsbA in periplasm.

Original languageEnglish
Pages (from-to)145-154
Number of pages10
JournalMicrobial Pathogenesis
Volume24
Issue number3
DOIs
Publication statusPublished - Mar 1998
Externally publishedYes

Fingerprint

Enterotoxins
Disulfides
Glutamic Acid
Hot Temperature
Escherichia coli
Periplasm
Amino Acids
Asparagine
Site-Directed Mutagenesis
Catalysis
Proline
Codon
Oligonucleotides
Oxidoreductases
Peptides

Keywords

  • Disulfide bond
  • Enterotoxin
  • Escherichia coli
  • Mutation
  • Oxireductase
  • Periplasm

ASJC Scopus subject areas

  • Microbiology
  • Infectious Diseases

Cite this

Involvement of glutamic acid residue at position 7 in the formation of the intramolecular disulfide bond of Escherichia coli heat-stable enterotoxin Ip in vivo. / Yamanaka, Hiroyasu; Nomura, Tomohiko; Okamoto, Keinosuke.

In: Microbial Pathogenesis, Vol. 24, No. 3, 03.1998, p. 145-154.

Research output: Contribution to journalArticle

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