Combination of arachidonic acid and guanosine 5'-O-(3-thiotriphosphate) induce translocation of rac p21s to membrane and activation of NADPH oxidase in a cell-free system

T Sawai; M Asada; H Nunoi; I Matsuda; S Ando; T Sasaki; K Kaibuchi; Y Takai; K Katayama

doi:10.1006/bbrc.1993.2039

Combination of arachidonic acid and guanosine 5'-O-(3-thiotriphosphate) induce translocation of rac p21s to membrane and activation of NADPH oxidase in a cell-free system

Biochem Biophys Res Commun. 1993 Aug 31;195(1):264-9. doi: 10.1006/bbrc.1993.2039.

Authors

T Sawai¹, M Asada, H Nunoi, I Matsuda, S Ando, T Sasaki, K Kaibuchi, Y Takai, K Katayama

Affiliation

¹ Department of Biochemistry, Tsukuba Research Laboratories, Eisai Co. Ltd., Ibaraki, Japan.

PMID: 8395827
DOI: 10.1006/bbrc.1993.2039

Abstract

The superoxide-generating NADPH oxidase system in phagocytes consists of membrane-associated cytochrome b558 and three cytosolic components named p67-phox, p47-phox, and rac p21s. In a cell-free system consisting of membrane and cytosol, the oxidase can be activated with guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S) and an unsaturated fatty acid such as arachidonic acid (AA). Incubation of cytosol and membrane with AA alone caused clear translocation of p47-phox and p67-phox to the membrane, but only slight translocation of rac p21s. GTP gamma S alone did not significantly induce the translocation of rac p21s. However, GTP gamma S in combination with AA markedly augmented rac p21s translocation to the membrane. The translocation of rac p21s is not induced by GDP or GDP beta S. These results indicate that the GTP-bound active form of rac p21s is the entity that is translocated to the membrane by the action of AA.

MeSH terms

Amino Acid Sequence
Arachidonic Acid / pharmacology*
Cell Differentiation / drug effects
Cell Membrane / metabolism
Cell-Free System
Cytosol / metabolism
Enzyme Activation
GTP-Binding Proteins / analysis
GTP-Binding Proteins / metabolism*
Guanosine 5'-O-(3-Thiotriphosphate) / pharmacology*
Guanosine Diphosphate / analogs & derivatives
Guanosine Diphosphate / pharmacology
Humans
Kinetics
Leukemia, Promyelocytic, Acute
Molecular Sequence Data
NADH, NADPH Oxidoreductases / metabolism*
NADPH Dehydrogenase / metabolism
NADPH Oxidases
Oligopeptides / chemical synthesis
Oligopeptides / immunology
Phosphoproteins / metabolism
Superoxides / metabolism
Thionucleotides / pharmacology
Tumor Cells, Cultured
rac GTP-Binding Proteins

Substances

Oligopeptides
Phosphoproteins
Thionucleotides
neutrophil cytosol factor 67K
Superoxides
Guanosine Diphosphate
Arachidonic Acid
Guanosine 5'-O-(3-Thiotriphosphate)
guanosine 5'-O-(2-thiodiphosphate)
NADH, NADPH Oxidoreductases
NADPH Oxidases
neutrophil cytosolic factor 1
NADPH Dehydrogenase
GTP-Binding Proteins
rac GTP-Binding Proteins