Identification of serine138 residue in the 4-residue segment k135k136i137s138 of luks-i component of staphylococcus intermedius leukocidin crucial for the luks-i-specific function of staphylococcal leukocidin

Akihito Nishiyama, Marie Antonette Ruth V. Guerra, Noriko Sugawara, Kenji Yokota, Jun Kaneko, Yoshiyuki Kamio

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5 Citations (Scopus)

Abstract

Luk-I produced by Staphylococcus intermedius was found to be a new member of the staphylococcal bi-component pore-forming toxin family, in which staphylococcal leukocidin, Panton-Valentine leukocidin, and γ-hemolysin are included. Luk-I consists of LukS-I and LukF-I. From the deduced amino acid sequence of LukS-I, a 4-residue sequence, K135K136I137S138, at the root of the stem region was found to be identical with that of the phosphorylated segment of a protein phosphorylated by protein kinase A. A mutant of LukS-I (MLSI-SA), in which the Ser138 residue was replaced by an alanine residue, was created, purified, and assayed for its leukocytolytic and pore-forming activities with LukF-I. Both LukS-I and MLSI-SA formed a ring-shaped complex with LukF-I on rabbit erythrocytes and human polymorphonuclear leukocytes (HPMNLs) membrane. However, MLSI-SA showed no leukocytolytic activity with LukF-I. LukS-I was phosphorylated by protein kinase A in the presence of [γ-32P] ATP in a cell-free system, but MLSI-SA was not phosphorylated significantly. A potent and selective inhibitor of protein kinase A (N- [2(p-bromocinnamylamino) ethyl]-5-isoquinolinesulfonamide (H-89)) showed 50% inhibition of the Luk-I-induced cell lysis at 0.5 nM. Thus, it is concluded that the phosphorylation of the Ser138 residue in the 4-residue segment K135K136I137S138 of LukS-I is important for the leukocytolysis of HPMNLS.

Original languageEnglish
Pages (from-to)328-335
Number of pages8
JournalBioscience, Biotechnology and Biochemistry
Volume66
Issue number2
DOIs
Publication statusPublished - 2002 Jan 1

Keywords

  • LukF-I
  • LukS-I
  • Protein kinase A inhibitor H-89
  • Staphylococcal leukocidin
  • Staphylococcus intermedius

ASJC Scopus subject areas

  • Biotechnology
  • Analytical Chemistry
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology
  • Organic Chemistry

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