Superoxide release and intracellular free calcium of calcium-depleted human neutrophils stimulated by N-formyl-methionyl-leucyl-phenylalanine

Miwako Nakagawara, Koichiro Takeshige, Hideki Sumimoto, Junichi Yoshitake, Skigeki Minakami

Research output: Contribution to journalArticle

44 Citations (Scopus)

Abstract

The superoxide release and the change in the intracellular free calcium ions on stimulation with N-formyl-methionyl-leucyl-phenylalanine were studied in human neutrophils deprived of divalent cations by treatment of the cells with ionophore A23187 in the presence of EGTA. The depleted cells showed no release of superoxide on stimulation with the chemotactic peptide when calcium ions were absent in the medium, but the activity was completely recovered when the cells were preincubated with calcium for at least 3 min before the stimulation. The cells pretreated with Cd2+ showed slight activity of the release, but no recovery was observed with other divalent cations such as Mg2+, Sr2+, Co2+, Ba2+ and Zn2+. The recovery with calcium ions was dependent on the time of the addition relative to the time of the stimulation with the chemotactic peptide: a simultaneous addition of both calcium and the peptide elicited about half of the full activity, while no release was observed when calcium was added later than 2 min after the stimulation with the peptide, though a marked elevation of the intracellular free calcium monitored by quin-2 fluorescence was found. Comparison of the time-courses of the superoxide release and the change in the fluorescence suggest that, besides the elevation of intracellular free calcium, a transient reaction which is also dependent on calcium is required for the full induction of the superoxide-producing activity.

Original languageEnglish
Pages (from-to)97-103
Number of pages7
JournalBBA - Molecular Cell Research
Volume805
Issue number1
DOIs
Publication statusPublished - Sep 14 1984

Fingerprint

N-Formylmethionine Leucyl-Phenylalanine
Superoxides
Neutrophils
Calcium
Peptides
Divalent Cations
Ions
Fluorescence
Egtazic Acid
Ionophores
Calcimycin

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Cell Biology
  • Molecular Biology

Cite this

Superoxide release and intracellular free calcium of calcium-depleted human neutrophils stimulated by N-formyl-methionyl-leucyl-phenylalanine. / Nakagawara, Miwako; Takeshige, Koichiro; Sumimoto, Hideki; Yoshitake, Junichi; Minakami, Skigeki.

In: BBA - Molecular Cell Research, Vol. 805, No. 1, 14.09.1984, p. 97-103.

Research output: Contribution to journalArticle

Nakagawara, Miwako ; Takeshige, Koichiro ; Sumimoto, Hideki ; Yoshitake, Junichi ; Minakami, Skigeki. / Superoxide release and intracellular free calcium of calcium-depleted human neutrophils stimulated by N-formyl-methionyl-leucyl-phenylalanine. In: BBA - Molecular Cell Research. 1984 ; Vol. 805, No. 1. pp. 97-103.
@article{bdea3aaa42b54dbd8d00f2e3c1b1277e,
title = "Superoxide release and intracellular free calcium of calcium-depleted human neutrophils stimulated by N-formyl-methionyl-leucyl-phenylalanine",
abstract = "The superoxide release and the change in the intracellular free calcium ions on stimulation with N-formyl-methionyl-leucyl-phenylalanine were studied in human neutrophils deprived of divalent cations by treatment of the cells with ionophore A23187 in the presence of EGTA. The depleted cells showed no release of superoxide on stimulation with the chemotactic peptide when calcium ions were absent in the medium, but the activity was completely recovered when the cells were preincubated with calcium for at least 3 min before the stimulation. The cells pretreated with Cd2+ showed slight activity of the release, but no recovery was observed with other divalent cations such as Mg2+, Sr2+, Co2+, Ba2+ and Zn2+. The recovery with calcium ions was dependent on the time of the addition relative to the time of the stimulation with the chemotactic peptide: a simultaneous addition of both calcium and the peptide elicited about half of the full activity, while no release was observed when calcium was added later than 2 min after the stimulation with the peptide, though a marked elevation of the intracellular free calcium monitored by quin-2 fluorescence was found. Comparison of the time-courses of the superoxide release and the change in the fluorescence suggest that, besides the elevation of intracellular free calcium, a transient reaction which is also dependent on calcium is required for the full induction of the superoxide-producing activity.",
author = "Miwako Nakagawara and Koichiro Takeshige and Hideki Sumimoto and Junichi Yoshitake and Skigeki Minakami",
year = "1984",
month = "9",
day = "14",
doi = "10.1016/0167-4889(84)90041-7",
language = "English",
volume = "805",
pages = "97--103",
journal = "Biochimica et Biophysica Acta - Molecular Cell Research",
issn = "0167-4889",
publisher = "Elsevier",
number = "1",

}

TY - JOUR

T1 - Superoxide release and intracellular free calcium of calcium-depleted human neutrophils stimulated by N-formyl-methionyl-leucyl-phenylalanine

AU - Nakagawara, Miwako

AU - Takeshige, Koichiro

AU - Sumimoto, Hideki

AU - Yoshitake, Junichi

AU - Minakami, Skigeki

PY - 1984/9/14

Y1 - 1984/9/14

N2 - The superoxide release and the change in the intracellular free calcium ions on stimulation with N-formyl-methionyl-leucyl-phenylalanine were studied in human neutrophils deprived of divalent cations by treatment of the cells with ionophore A23187 in the presence of EGTA. The depleted cells showed no release of superoxide on stimulation with the chemotactic peptide when calcium ions were absent in the medium, but the activity was completely recovered when the cells were preincubated with calcium for at least 3 min before the stimulation. The cells pretreated with Cd2+ showed slight activity of the release, but no recovery was observed with other divalent cations such as Mg2+, Sr2+, Co2+, Ba2+ and Zn2+. The recovery with calcium ions was dependent on the time of the addition relative to the time of the stimulation with the chemotactic peptide: a simultaneous addition of both calcium and the peptide elicited about half of the full activity, while no release was observed when calcium was added later than 2 min after the stimulation with the peptide, though a marked elevation of the intracellular free calcium monitored by quin-2 fluorescence was found. Comparison of the time-courses of the superoxide release and the change in the fluorescence suggest that, besides the elevation of intracellular free calcium, a transient reaction which is also dependent on calcium is required for the full induction of the superoxide-producing activity.

AB - The superoxide release and the change in the intracellular free calcium ions on stimulation with N-formyl-methionyl-leucyl-phenylalanine were studied in human neutrophils deprived of divalent cations by treatment of the cells with ionophore A23187 in the presence of EGTA. The depleted cells showed no release of superoxide on stimulation with the chemotactic peptide when calcium ions were absent in the medium, but the activity was completely recovered when the cells were preincubated with calcium for at least 3 min before the stimulation. The cells pretreated with Cd2+ showed slight activity of the release, but no recovery was observed with other divalent cations such as Mg2+, Sr2+, Co2+, Ba2+ and Zn2+. The recovery with calcium ions was dependent on the time of the addition relative to the time of the stimulation with the chemotactic peptide: a simultaneous addition of both calcium and the peptide elicited about half of the full activity, while no release was observed when calcium was added later than 2 min after the stimulation with the peptide, though a marked elevation of the intracellular free calcium monitored by quin-2 fluorescence was found. Comparison of the time-courses of the superoxide release and the change in the fluorescence suggest that, besides the elevation of intracellular free calcium, a transient reaction which is also dependent on calcium is required for the full induction of the superoxide-producing activity.

UR - http://www.scopus.com/inward/record.url?scp=0021166775&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0021166775&partnerID=8YFLogxK

U2 - 10.1016/0167-4889(84)90041-7

DO - 10.1016/0167-4889(84)90041-7

M3 - Article

VL - 805

SP - 97

EP - 103

JO - Biochimica et Biophysica Acta - Molecular Cell Research

JF - Biochimica et Biophysica Acta - Molecular Cell Research

SN - 0167-4889

IS - 1

ER -