PMID- 2164554
OWN - NLM
STAT- MEDLINE
DCOM- 19900820
LR  - 20190516
IS  - 0741-5400 (Print)
IS  - 0741-5400 (Linking)
VI  - 48
IP  - 2
DP  - 1990 Aug
TI  - Activation of the superoxide-generating NADPH oxidase of macrophages by sodium 
      dodecyl sulfate in a soluble cell-free system: evidence for involvement of a G 
      protein.
PG  - 107-15
AB  - The superoxide (O2-)-generating NADPH oxidase of resting macrophages can be 
      activated in a soluble cell-free system by certain anionic amphiphiles, such as 
      sodium dodecyl sulfate (SDS). We demonstrate that cell-free activation is 
      specifically enhanced by nonhydrolyzable guanosine 5'-triphosphate (GTP) 
      analogues. Guanosine 5'-diphosphate (GDP) and guanosine 5'-O-(2-thiodiphosphate) 
      (GDP beta S) prevent cell-free oxidase activation and reverse the activated state 
      when added to preactivated oxidase preparations. Nonhydrolyzable GTP analogues 
      have a protective effect on the SDS-stimulated NADPH oxidase, as shown by the 
      maintenance of more than 90% and close to 60% of enzyme activity 6 and 24 hr, 
      respectively, after the addition of SDS to the cell-free preparation. A novel 
      procedure is described for separating the activated NADPH oxidase from SDS and 
      added nucleotides by gel filtration of the SDS-stimulated solubilized 
      membrane-cytosol mixtures through a Sephadex G-25 column. By utilizing this 
      method, it was found that the presence of micromolar concentrations of 
      nonhydrolyzable GTP analogues during activation by SDS results in a marked 
      increase in the recovery of SDS-independent, NADPH-dependent O2(-)-producing 
      activity in the excluded volume of the column. It is suggested that GTP 
      stabilizes the SDS-induced complex between membrane and cytosolic components of 
      the oxidase. Cell-free activation of NADPH oxidase by SDS was found to be cholera 
      and pertussis toxin insensitive. These results serve as evidence of the 
      participation of a G protein in the activation of the O2(-)-generating NADPH 
      oxidase of macrophages by SDS.
FAU - Aharoni, I
AU  - Aharoni I
AD  - Department of Human Microbiology, Sackler School of Medicine, Tel Aviv 
      University, Israel.
FAU - Pick, E
AU  - Pick E
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - J Leukoc Biol
JT  - Journal of leukocyte biology
JID - 8405628
RN  - 0 (Thionucleotides)
RN  - 0 (Virulence Factors, Bordetella)
RN  - 11062-77-4 (Superoxides)
RN  - 146-91-8 (Guanosine Diphosphate)
RN  - 368GB5141J (Sodium Dodecyl Sulfate)
RN  - 37589-80-3 (Guanosine 5'-O-(3-Thiotriphosphate))
RN  - 86-01-1 (Guanosine Triphosphate)
RN  - 9012-63-9 (Cholera Toxin)
RN  - EC 1.6.- (NADH, NADPH Oxidoreductases)
RN  - EC 1.6.3.- (NADPH Oxidases)
RN  - EC 2.4.2.31 (Pertussis Toxin)
RN  - EC 3.6.1.- (GTP-Binding Proteins)
SB  - IM
MH  - Animals
MH  - Cell-Free System
MH  - Cholera Toxin/pharmacology
MH  - Enzyme Activation
MH  - GTP-Binding Proteins/*physiology
MH  - Guanosine 5'-O-(3-Thiotriphosphate)
MH  - Guanosine Diphosphate/pharmacology
MH  - Guanosine Triphosphate/analogs & derivatives/pharmacology
MH  - Guinea Pigs
MH  - Macrophages/*enzymology
MH  - NADH, NADPH Oxidoreductases/*analysis/antagonists & inhibitors
MH  - NADPH Oxidases
MH  - Pertussis Toxin
MH  - Sodium Dodecyl Sulfate/*pharmacology
MH  - Superoxides/*metabolism
MH  - Thionucleotides/pharmacology
MH  - Virulence Factors, Bordetella/pharmacology
EDAT- 1990/08/01 00:00
MHDA- 1990/08/01 00:01
CRDT- 1990/08/01 00:00
PHST- 1990/08/01 00:00 [pubmed]
PHST- 1990/08/01 00:01 [medline]
PHST- 1990/08/01 00:00 [entrez]
AID - 10.1002/jlb.48.2.107 [doi]
PST - ppublish
SO  - J Leukoc Biol. 1990 Aug;48(2):107-15. doi: 10.1002/jlb.48.2.107.