Role of Mg 2+ in activation of nadph oxidase of human neutrophils

Evidence that Mg 2+ acts through G-protein

Kunihiko Aoyagi, Koichiro Takeshige, Hideki Sumimoto, Hiroyuki Nunoi, Shigeki Minakami

Research output: Contribution to journalArticle

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Abstract

The membrane fraction and three cytosolic proteins of neutrophils, p47-phox, p67-phox and a G-protein, are involved in the cell-free activation of the O 2 - -generating NADPH oxidase in the presence of SDS, though it has been controversial whether the G-protein is required or just enhancing the activity. We have used the three cytosolic factors, the solubilized membrane fraction, GTPγS and SDS, and found that both G-protein and GTPγS are essential for the activation of the NADPH oxidase. The effect of GTPγS is modified by Mg 2+ : the cations enhance the O 2 - generation at low concentrations of GTPγS, whereas they attenuate the activity at higher concentrations of GTPγS. In the presence of 10 μM GTPγS, the maximal activity is observed at 0.1 μM Mg 2+ , which is several-fold higher than that at 1 mM Mg 2+ . The omission of Mg 2+ followed by the chelation with EDTA results in loss of the activation, which is completely restored by the addition of Mg 2+ . Thus, Mg 2+ seems to modulate the activation of the NADPH oxidase at the level of the G-protein.

Original languageEnglish
Pages (from-to)391-397
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume186
Issue number1
DOIs
Publication statusPublished - Jul 15 1992

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GTP-Binding Proteins
NADPH Oxidase
Oxidoreductases
Neutrophils
Chemical activation
Membranes
Chelation
Edetic Acid
Cations
Proteins

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Role of Mg 2+ in activation of nadph oxidase of human neutrophils : Evidence that Mg 2+ acts through G-protein. / Aoyagi, Kunihiko; Takeshige, Koichiro; Sumimoto, Hideki; Nunoi, Hiroyuki; Minakami, Shigeki.

In: Biochemical and Biophysical Research Communications, Vol. 186, No. 1, 15.07.1992, p. 391-397.

Research output: Contribution to journalArticle

Aoyagi, Kunihiko ; Takeshige, Koichiro ; Sumimoto, Hideki ; Nunoi, Hiroyuki ; Minakami, Shigeki. / Role of Mg 2+ in activation of nadph oxidase of human neutrophils : Evidence that Mg 2+ acts through G-protein. In: Biochemical and Biophysical Research Communications. 1992 ; Vol. 186, No. 1. pp. 391-397.
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