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
N1 - Funding Information:
This study was supportedin part by grants from the Ministry of EducationS, ciencea nd Culture and the Ministry of Health and Welfare, Japan.
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.
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U2 - 10.1016/0167-4889(84)90041-7
DO - 10.1016/0167-4889(84)90041-7
M3 - Article
C2 - 6089914
AN - SCOPUS:0021166775
SN - 0167-4889
VL - 805
SP - 97
EP - 103
JO - BBA - Molecular Cell Research
JF - BBA - Molecular Cell Research
IS - 1
ER -