A prodigiosin analogue inactivates NADPH oxidase in macrophage cells by inhibiting assembly of p47phox and Rac

Takuji Nakashima, Takashi Iwashita, Tsuyoshi Fujita, Emiko Sato, Yoshimi Niwano, Masahiro Kohno, Shunsuke Kuwahara, Nobuyuki Harada, Satoshi Takeshita, Tatsuya Oda

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Prodigiosins are natural red pigments that have multi-biological activities. Recently, we discovered a marine bacterial strain, which produces a red pigment. Extensive two-dimensional nuclear magnetic resonance and mass spectrometry analysis showed that the pigment is a prodigiosin analogue (PG-L-1). Here, we investigated the effect of PG-L-1 on NADPH oxidase activity in macrophage cells. PG-L-1 significantly inhibited superoxide anion (O 2-) production by phorbol 12-myristate 13-acetate (PMA)-stimulated RAW 264.7 cells, a mouse macrophage cell line. The ED 50 value was estimated to be ∼0.3 μM. PG-L-1 had no direct scavenging effect on O2- generated by hypoxanthine/ xanthine oxidase system in electron spin resonance-spin trapping determinations, suggesting that this compound directly acts on the O2- production system, NADPH oxidase, in macrophage cells. We further investigated the effect of PG-L-1 on the behaviour of the cytosolic components of the NADPH oxidase, p67phox, p47phox, p40phox, Rac and protein kinase C (PKC), in PMA-stimulated RAW 264.7 cells. Although PG-L-1 showed no effect on the activation of PKC, the immunoblotting analysis using specific antibodies showed that PG-L-1 strongly inhibits the association of p47phox and Rac in the plasma membrane of PMA-stimulated RAW 264.7 cells. These results suggest that PG-L-1 inactivates NADPH oxidase through the inhibition of the binding of p47phox and Rac to the membrane components of NADPH oxidase.

Original languageEnglish
Pages (from-to)107-115
Number of pages9
JournalJournal of biochemistry
Volume143
Issue number1
DOIs
Publication statusPublished - 2008 Jan 1

Keywords

  • NADPH oxidase inhibitor
  • Prodigiosin
  • Rac protein
  • Superoxide
  • p47phox

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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