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Activation of the respiratory burst enzyme in human polymorphonuclear leukocytes by chemoattractants and other soluble stimuli. Evidence that the same oxidase is activated by different transductional mechanisms.

Publication ,  Journal Article
McPhail, LC; Snyderman, R
Published in: J Clin Invest
July 1983

Chemoattractant-receptor coupling triggers several biologic responses in phagocytic cells including activation of the respiratory burst. Prior evidence in intact cells implied that stimulation of the respiratory burst by chemoattractants was by a mechanism different from other soluble agents suggesting the possibility that different oxidative enzymes were responsible. We now show that the chemoattractants N-formyl-methionyl-leucyl-phenylalanine and a split fragment of the fifth component of complement (C5a) stimulate an NADPH oxidase activity, measured in the 50,000-g particulate fraction from human polymorphonuclear leukocytes (PMN). Levels of oxidase activity stimulated by the chemoattractants were both time and dose dependent and required the presence of cytochalasin B during stimulation. In contrast, activation by two nonchemotactic stimuli, the ionophore A23187 and phorbol myristate acetate (PMA), did not require cytochalasin B. Temporal patterns of oxidase activation suggested that different stimuli follow different transductional pathways. Chemoattractant-mediated activation was immediate (no lag); peaked by 45 s and declined rapidly to approximately 50% of maximal by 2 min. In contrast, activation by A23187 or PMA had a 15-30-s lag and increased more slowly. Stimulation by A23187 peaked at 5 min, then declined. Stimulation by PMA plateaued at 20 min and did not decline by 90 min. Comparison of Km values for NADPH and NADH obtained by Lineweaver-Burk analysis of the oxidase activity stimulated by N-formyl-methionyl-leucyl-phenylalanine, A23187, and PMA suggested that the same enzyme was activated by all stimuli. Thus, chemoattractants and other soluble stimuli appear to activate the same respiratory burst enzyme in PMN but they utilize different transductional mechanisms and are regulated differently.

Duke Scholars

Published In

J Clin Invest

DOI

ISSN

0021-9738

Publication Date

July 1983

Volume

72

Issue

1

Start / End Page

192 / 200

Location

United States

Related Subject Headings

  • Tetradecanoylphorbol Acetate
  • Oligopeptides
  • Neutrophils
  • NADPH Oxidases
  • NADH, NADPH Oxidoreductases
  • N-Formylmethionine Leucyl-Phenylalanine
  • N-Formylmethionine
  • Methionine
  • Kinetics
  • Immunology
 

Citation

APA
Chicago
ICMJE
MLA
NLM

Published In

J Clin Invest

DOI

ISSN

0021-9738

Publication Date

July 1983

Volume

72

Issue

1

Start / End Page

192 / 200

Location

United States

Related Subject Headings

  • Tetradecanoylphorbol Acetate
  • Oligopeptides
  • Neutrophils
  • NADPH Oxidases
  • NADH, NADPH Oxidoreductases
  • N-Formylmethionine Leucyl-Phenylalanine
  • N-Formylmethionine
  • Methionine
  • Kinetics
  • Immunology