Activation of the neutrophil respiratory burst requires both intracellular and extracellular calcium

W. K. Kim-Park, M. A. Moore, Z. W. Hakki, Michael Kowolik

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

Activation of neutrophil oxidases, including NADPH oxidase, is Ca2+ dependent. The aim of this study was to determine the roles of intra- and extracellular Ca2+, leading to generation of the respiratory burst, as monitored by luminol-dependent chemiluminescence (CL). All results were recorded as integrals (millivolt.min) and compared by a two-tail Student's t test. Preincubation of cells with chelators of intra- or extracellular Ca2+ inhibited N-Formyl-Met-Leu-Phe (FMLP)-stimulated burst activity (p <0.01). In contrast, stimulation by phorbol myristate acetate (PMA), while inhibited by extracellular Ca2+ chelation with EGTA (p <0.001), was potentiated by intracellular Ca2+ chelation with BAPTA (p <0.01). This suggests that the protein kinase C (PKC)-mediated burst may be diminished by intracellular Ca2+-dependent phosphatase. A selective inhibitor of tyrosine phosphatase, sodium vanadate, potentiated CL generation by both FMLP and PMA, indicating a dominant phosphatase activation with transiently increased Ca2+, masking the kinase-mediated respiratory burst. The selective inhibitors of PKC or tyrosine kinase prevented PMA and vanadate/PMA stimulation (p <0.005). Furthermore, the putative Ca2+ channel agonists glutamate (10-5M) and N-methyl-D-aspartate (NMDA) (10-5M) alone failed to influence CL output, but produced marked potentiation following pre-treatment with vanadate. Again this indicates a dominant activation of phosphatase triggered by the glutamate-mediated Ca2+ influx, so masking the kinase-dependent NADPH oxidase activity. A competitive antagonist of NMDA, AP7, significantly decreased vanadate-mediated CL in an EGTA-sensitive manner (p <0.001). The data confirm a requirement for intra- and extracellular Ca2+ in neutrophil respiratory burst activation via the kinase/phosphatase cycle, and an agonist effect by NMDA within the Ca2+ cascade mechanism.

Original languageEnglish (US)
Pages (from-to)394-404
Number of pages11
JournalAnnals of the New York Academy of Sciences
Volume832
DOIs
StatePublished - Dec 15 1997

Fingerprint

Neutrophil Activation
Respiratory Burst
Phosphoric Monoester Hydrolases
Vanadates
Chemiluminescence
Tetradecanoylphorbol Acetate
Luminescence
Chemical activation
Calcium
N-Methylaspartate
2-amino-7-phosphonoheptanoic acid
Phosphotransferases
NADPH Oxidase
Egtazic Acid
Chelation
methionyl-leucyl-phenylalanine
Excitatory Amino Acid Agonists
Luminol
Chelating Agents
Protein-Tyrosine Kinases

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Activation of the neutrophil respiratory burst requires both intracellular and extracellular calcium. / Kim-Park, W. K.; Moore, M. A.; Hakki, Z. W.; Kowolik, Michael.

In: Annals of the New York Academy of Sciences, Vol. 832, 15.12.1997, p. 394-404.

Research output: Contribution to journalArticle

@article{5b59b6b8b49843499c155751e7993048,
title = "Activation of the neutrophil respiratory burst requires both intracellular and extracellular calcium",
abstract = "Activation of neutrophil oxidases, including NADPH oxidase, is Ca2+ dependent. The aim of this study was to determine the roles of intra- and extracellular Ca2+, leading to generation of the respiratory burst, as monitored by luminol-dependent chemiluminescence (CL). All results were recorded as integrals (millivolt.min) and compared by a two-tail Student's t test. Preincubation of cells with chelators of intra- or extracellular Ca2+ inhibited N-Formyl-Met-Leu-Phe (FMLP)-stimulated burst activity (p <0.01). In contrast, stimulation by phorbol myristate acetate (PMA), while inhibited by extracellular Ca2+ chelation with EGTA (p <0.001), was potentiated by intracellular Ca2+ chelation with BAPTA (p <0.01). This suggests that the protein kinase C (PKC)-mediated burst may be diminished by intracellular Ca2+-dependent phosphatase. A selective inhibitor of tyrosine phosphatase, sodium vanadate, potentiated CL generation by both FMLP and PMA, indicating a dominant phosphatase activation with transiently increased Ca2+, masking the kinase-mediated respiratory burst. The selective inhibitors of PKC or tyrosine kinase prevented PMA and vanadate/PMA stimulation (p <0.005). Furthermore, the putative Ca2+ channel agonists glutamate (10-5M) and N-methyl-D-aspartate (NMDA) (10-5M) alone failed to influence CL output, but produced marked potentiation following pre-treatment with vanadate. Again this indicates a dominant activation of phosphatase triggered by the glutamate-mediated Ca2+ influx, so masking the kinase-dependent NADPH oxidase activity. A competitive antagonist of NMDA, AP7, significantly decreased vanadate-mediated CL in an EGTA-sensitive manner (p <0.001). The data confirm a requirement for intra- and extracellular Ca2+ in neutrophil respiratory burst activation via the kinase/phosphatase cycle, and an agonist effect by NMDA within the Ca2+ cascade mechanism.",
author = "Kim-Park, {W. K.} and Moore, {M. A.} and Hakki, {Z. W.} and Michael Kowolik",
year = "1997",
month = "12",
day = "15",
doi = "10.1111/j.1749-6632.1997.tb46267.x",
language = "English (US)",
volume = "832",
pages = "394--404",
journal = "Annals of the New York Academy of Sciences",
issn = "0077-8923",
publisher = "Wiley-Blackwell",

}

TY - JOUR

T1 - Activation of the neutrophil respiratory burst requires both intracellular and extracellular calcium

AU - Kim-Park, W. K.

AU - Moore, M. A.

AU - Hakki, Z. W.

AU - Kowolik, Michael

PY - 1997/12/15

Y1 - 1997/12/15

N2 - Activation of neutrophil oxidases, including NADPH oxidase, is Ca2+ dependent. The aim of this study was to determine the roles of intra- and extracellular Ca2+, leading to generation of the respiratory burst, as monitored by luminol-dependent chemiluminescence (CL). All results were recorded as integrals (millivolt.min) and compared by a two-tail Student's t test. Preincubation of cells with chelators of intra- or extracellular Ca2+ inhibited N-Formyl-Met-Leu-Phe (FMLP)-stimulated burst activity (p <0.01). In contrast, stimulation by phorbol myristate acetate (PMA), while inhibited by extracellular Ca2+ chelation with EGTA (p <0.001), was potentiated by intracellular Ca2+ chelation with BAPTA (p <0.01). This suggests that the protein kinase C (PKC)-mediated burst may be diminished by intracellular Ca2+-dependent phosphatase. A selective inhibitor of tyrosine phosphatase, sodium vanadate, potentiated CL generation by both FMLP and PMA, indicating a dominant phosphatase activation with transiently increased Ca2+, masking the kinase-mediated respiratory burst. The selective inhibitors of PKC or tyrosine kinase prevented PMA and vanadate/PMA stimulation (p <0.005). Furthermore, the putative Ca2+ channel agonists glutamate (10-5M) and N-methyl-D-aspartate (NMDA) (10-5M) alone failed to influence CL output, but produced marked potentiation following pre-treatment with vanadate. Again this indicates a dominant activation of phosphatase triggered by the glutamate-mediated Ca2+ influx, so masking the kinase-dependent NADPH oxidase activity. A competitive antagonist of NMDA, AP7, significantly decreased vanadate-mediated CL in an EGTA-sensitive manner (p <0.001). The data confirm a requirement for intra- and extracellular Ca2+ in neutrophil respiratory burst activation via the kinase/phosphatase cycle, and an agonist effect by NMDA within the Ca2+ cascade mechanism.

AB - Activation of neutrophil oxidases, including NADPH oxidase, is Ca2+ dependent. The aim of this study was to determine the roles of intra- and extracellular Ca2+, leading to generation of the respiratory burst, as monitored by luminol-dependent chemiluminescence (CL). All results were recorded as integrals (millivolt.min) and compared by a two-tail Student's t test. Preincubation of cells with chelators of intra- or extracellular Ca2+ inhibited N-Formyl-Met-Leu-Phe (FMLP)-stimulated burst activity (p <0.01). In contrast, stimulation by phorbol myristate acetate (PMA), while inhibited by extracellular Ca2+ chelation with EGTA (p <0.001), was potentiated by intracellular Ca2+ chelation with BAPTA (p <0.01). This suggests that the protein kinase C (PKC)-mediated burst may be diminished by intracellular Ca2+-dependent phosphatase. A selective inhibitor of tyrosine phosphatase, sodium vanadate, potentiated CL generation by both FMLP and PMA, indicating a dominant phosphatase activation with transiently increased Ca2+, masking the kinase-mediated respiratory burst. The selective inhibitors of PKC or tyrosine kinase prevented PMA and vanadate/PMA stimulation (p <0.005). Furthermore, the putative Ca2+ channel agonists glutamate (10-5M) and N-methyl-D-aspartate (NMDA) (10-5M) alone failed to influence CL output, but produced marked potentiation following pre-treatment with vanadate. Again this indicates a dominant activation of phosphatase triggered by the glutamate-mediated Ca2+ influx, so masking the kinase-dependent NADPH oxidase activity. A competitive antagonist of NMDA, AP7, significantly decreased vanadate-mediated CL in an EGTA-sensitive manner (p <0.001). The data confirm a requirement for intra- and extracellular Ca2+ in neutrophil respiratory burst activation via the kinase/phosphatase cycle, and an agonist effect by NMDA within the Ca2+ cascade mechanism.

UR - http://www.scopus.com/inward/record.url?scp=0031574408&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0031574408&partnerID=8YFLogxK

U2 - 10.1111/j.1749-6632.1997.tb46267.x

DO - 10.1111/j.1749-6632.1997.tb46267.x

M3 - Article

C2 - 9704067

AN - SCOPUS:0031574408

VL - 832

SP - 394

EP - 404

JO - Annals of the New York Academy of Sciences

JF - Annals of the New York Academy of Sciences

SN - 0077-8923

ER -