The GDP-bound form of the small G protein Rac1 p21 is a potent activator of the superoxide-forming NADPH oxidase of macrophages.
Phagocytes produce superoxide by the assembly of a multicomponent complex that utilizes NADPH for the reduction of molecular oxygen (NADPH oxidase). The components participating in the assembly are a membrane-bound flavocytochrome and three cytosolic proteins, one of which was shown to be a dimer of the small GTP-binding protein (G protein) Rac1 p21 or Rac2 p21 with GDP dissociation inhibitor for Rho (Rho GDI). We determined the identity and quantity of the nucleotide bound to Rac1 p21 by high performance anion exchange chromatography of extracts prepared from highly purified Rac1 p21-Rho GDI, isolated from guinea pig macrophage cytosol. Rac1 p21 contained only GDP at a ratio of close to 1 mol of GDP per mol of G protein. The GDP-bound form of Rac1 p21 complexed to Rho GDI functioned as a potent activator of NADPH oxidase in a cell-free system that contained no free GTP or ATP. We propose that the GDP-bound form of Rac1 p21 might be the physiological activator of NADPH oxidase in macrophages, following its dissociation from Rho GDI, and that nucleotide exchange or conversion to GTP is not necessarily involved.
Bromberg
Y
Y
Department of Human Microbiology, Sackler School of Medicine, Tel Aviv University, Israel.
Shani
E
E
Joseph
G
G
Gorzalczany
Y
Y
Sperling
O
O
Pick
E
E
D000595:Amino Acid Sequence; D000818:Animals; D004789:Enzyme Activation; D019204:GTP-Binding Proteins / Q000378:metabolism*; D006153:Guanosine Diphosphate / Q000378:metabolism*; D006168:Guinea Pigs; D008264:Macrophages / Q000201:enzymology*; D008969:Molecular Sequence Data; D009247:NADH, NADPH Oxidoreductases / Q000378:metabolism*; D019255:NADPH Oxidases; D009711:Nucleotides / Q000378:metabolism; D011485:Protein Binding; D013481:Superoxides / Q000378:metabolism*; D020744:rac GTP-Binding Proteins
D009711:Nucleotides; D013481:Superoxides; D006153:Guanosine Diphosphate; D009247:NADH, NADPH Oxidoreductases; D019255:NADPH Oxidases; D019204:GTP-Binding Proteins; D020744:rac GTP-Binding Proteins