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PMID: 2831270 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Certain lymphoid cells contain the membrane-associated component of the phagocyte-specific NADPH oxidase.

Journal of immunology (Baltimore, Md. : 1950) ·Vol. 140 ·No. 5 ·1988-03-01 ·Pages 1611-7

Pick E, Gadba R

Abstract

Anionic amphiphiles such as long chain unsaturated fatty acids and SDS were shown to activate the superoxide (O2-) producing NADPH oxidase in a cell-free system derived from sonically disrupted phagocytes (macrophages and granulocytes). O2- production required the cooperation of a membrane associated component sedimenting at 48,000 X g (pi) and a cytosolic factor (sigma). The purpose of the present investigation was to find out whether components pi and sigma were also present in non-phagocytic cells that do not produce O2- when stimulated. It was found that the 48,000 X g pellets of guinea pig lymph node and thymus cell sonicates contained significant amounts of component pi, as shown by their ability to support SDS-elicited NADPH-dependent O2- production when supplemented with macrophage cytosol. Lymph node and thymus pi could be extracted from the membrane by 30 mM octyl glucoside, just as its macrophage-derived equivalent. Combining lymph node and thymus 48,000 X g pellet with autologous cytosol did not yield an active enzyme preparation. Also, cytosol from lymph node and thymus cells could not cooperate with macrophage 48,000 X g pellet, indicating that component sigma was lacking in lymphoid cells. Neither pi nor sigma could be detected in guinea pig kidney, the mouse myeloma cell line MOPC 315 and the canine cell line Cf2Th. The 48,000 X g pellet of all nonphagocytic cells examined contained a b-cytochrome that resembled, by its spectral characteristics, the cytochrome b559 thought to be characteristic of phagocytes. In macrophages, cytochrome b559 represented 80% of b-cytochrome content of the 48,000 X g pellet, whereas in non-phagocytic cells, the equivalent material represented only 50 to 60%. There was no correlation between the presence and quantity of the cytochrome b559-like chromophore in the 48,000 X g pellet of a particular cell type and its ability to cooperate with macrophage cytosol in SDS-elicited O2- production.

MeSH Terms
Animals Cell Line Cell Membrane/enzymology,metabolism Cytochrome b Group/isolation & purification,metabolism Dogs Guinea Pigs Lymphocytes/analysis,enzymology,metabolism Mice NADH, NADPH Oxidoreductases/isolation & purification,metabolism Phagocytes/enzymology,metabolism Photosystem II Protein Complex Subcellular Fractions/metabolism Superoxides/biosynthesis
Chemicals
Cytochrome b Group Photosystem II Protein Complex Superoxides cytochrome b559 NADH, NADPH Oxidoreductases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Pick E
Department of Human Microbiology, Sackler School of Medicine, Tel-Aviv, Israel.
Gadba R
Article Info
Journal
Journal of immunology (Baltimore, Md. : 1950)
Abbr.
J Immunol
ISSN
0022-1767
Published
1988-03-01
Pages
1611-7
Language
English
Region
United States
NLM ID
2985117R
Subset
IM
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