Pyridine nucleotide-dependent superoxide production by a cell-free system from human granulocytes.
Open accessAbstract
Using an assay that measured superoxide dismutase-inhibitable nitro blue tetrazolium reduction, we studied superoxide (O2-) production by a cell-free system from human granulocytes. At 40 muM NADPH and a protein concentration of 0.12 mg/ml, lysates prepared from human granulocytes formed O2- at a rate of 18. 4 +/- 4.6 SE nmol/ml reaction mixture per donor, but not with glucose-6-phosphate, 6-phosphogluconate, glyceraldehyde-3-phosphate, sodium lactate, glutathione, or ascorbic acid. The Km's for NADPH and NADH were 8.6 +/- 4.6 muM and 0.83 +/- 0.30 mM, respectively, suggesting that NADPH is the physiological electron donor in this system. O2- production was not inhibited by 1mM KCN. The rate of O2- production by the cell-free system was comparable to the rate of O2-production by an equivalent quantity of intact granulocytes incubated under similar conditions. O2- production by lysates from granulocytes preincubated with serum under conditions previously shown to stimulate O2- production in the intact cells was no different than its production by lysates from unstimulated cells. O2- production at 0.2 mM and 0.02 mM NADPH by lysates from the granulocytes of two patients with chronic granulomatous disease was similar to O2- production by control lysates. This finding was interpreted in terms of the possibility that the metabolic lesion in chronic granulomatous disease may lie outside the oxygen-metabolizing enzyme system of the granulocyte, or alternatively, that the granulocytes may contain two O2- - forming enzymes, one of which is inactive in chronic granulomatous disease.
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NADPH is the preferred substrate for all NOX isoforms, with the NOX2 isoform being particularly NADPH specific.
“NADPH is the preferred substrate for all NOX isoforms, with the NOX2 isoform being particularly NADPH specific (48, 63, 177, 1047).”
From Sources of Vascular Nitric Oxide and Reactive Oxygen Species and Their Regulation (2019) · cites this paper as [48]
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- Authors
- Babior, B. M., Curnutte, J. T., & Kipnes, B. S.
- Affiliations
- Tufts Medical Center (US)
- Venue
- Journal of Clinical Investigation
- Published
- 1975-10-01
- Volume
- 56
- Issue
- 4
- Pages
- 1035-1042
- ISSN
- 0021-9738 (ISSN-L)
- PMC
- PMC301959
- OpenAlex
- W2166083878
- Cited by
- 112
- References
- 27
- Citations by year
- 2024: 2 · 2023: 2 · 2022: 1 · 2021: 3 · 2020: 4 · 2019: 1 · 2018: 3 · 2017: 3 · 2016: 2 · 2014: 4 · 2013: 1 · 2012: 1
- FWCI
- 2.51
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Journal of Clinical Investigation (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- U.S. Army Medical Research and Development Command
- Topics
- Life Sciences › Immunology and Microbiology › Immunology › Neutrophil, Myeloperoxidase and Oxidative Mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Redox biology and oxidative stress
- Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Toxicology › Bioactive Compounds and Antitumor Agents
- Keywords
- superoxide production, human granulocytes, NADPH, NADH, cell-free system, chronic granulomatous disease, superoxide dismutase, nitroblue tetrazolium reduction, oxygen metabolism, serum stimulation
- MeSH terms
- Cell-Free System, Cytochrome c Group, Granulocytes, Granulomatous Disease, Chronic, Humans, Leukocytes, NAD, NADP, Oxidation-Reduction, Oxygen, Oxygen Consumption, Proteins, Superoxides
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