Summary
Professional phagocytes, neutrophils, possess a unique membrane-associated NADPH-oxidase system, dormant in resting cells, which becomes activated upon exposure to the appropriate stimuli and catalyzes the one-electron reduction of molecular oxygen to superoxide, O −2 . Oxidase activation involves the assembly, in the plasma membrane, of membrane-bound and cytosolic constituents of the oxidase system, which are disassembled in the resting state. The oxidase system consists of two plasma membrane-bound components; low-potential cytochromeb 558, which is composed of two subunits of 22-kDa, and 91-kDa, and a possible flavoprotein related to the electron transport between NADPH and cytochromeb 558. Recent reports have indicated that FAD-binding sites of the oxidase are contained in cytochromeb 558. At least two cytosolic components, 67-kDa protein and a phosphorylated 47-kDa protein, are known to translocate to the plasma membrane, ensuring assembly of an active O −2 -generating NADPH-oxidase system. It is the purpose of this review to focus on recent data concerning electron transfer mechanisms of the activated neutrophil NADPH-oxidase complex and molecular pathology of chronic granulomatous disease.
Similar content being viewed by others
References
Segal AW (1989) The electron transport chain of the microbicidal oxidase of phagocytic cells and its involvement in the molecular pathology of chronic granulomatous disease. J Clin Invest 83:1785–1793
Cross AR, Jones OTJ (1991) Enzymic mechanisms of superoxide production. Biochim Biophys Acta 1057:281–298
Morel F, Doussière J, Vignais PV (1991) The superoxidegenerating oxidase of phagocytic cells: physiological, molecular and pathological aspects. Eur J Biochem 201:523–546
Babior BM, Kipnes RS, Curnutte JT (1973) Biological defense mechanisms. The production by leukocytes of superoxide, a potential bactericidal agent. J Clin Invest 52:741–744
Babior BM (1978) Oxygen-dependent microbial killing by phagocytes. N Engl J Med 298:659–668
Rossi F, Romeo D, Patriarca P (1972) Mechanism of phagocytosis-associated oxidative metabolism in polymorphonuclear leukocytes and macrophages. J Reticuloendothel Soc 12:127–149
Repine JE, White JH, Clawson CC, Holmes BM (1974) The influence of phorbol myristate acetate on oxygen consumption by polymorphonuclear leukocytes. J Lab Clin Med 83:911–920
Smith RM, Curnutte JR (1991) Molecular basis of chronic granulomatous disease. Blood 77:673–686
Segal AW, Jones OTG (1980) Rapid incorporation of the human neutrophil plasma membrane cytochromeb into phagocytic vacuoles. Biochem Biophys Res Commun 92:710–715
Garcia RC, Segal AW (1984) Changes in the subcellular distribution of the cytochromeb-245 on stimulation of human neutrophils. Biochem J 219:233–242
Segal AW, Jones OTG (1978) Novel cytochromeb system in phagocytic vacuoles from human granulocytes. Nature 276:515–517
Cross AR, Jones OTG, Harper AM, Segal AW (1981) Oxidation-reduction properties of the cytochromeb found in the plasma-membrane fraction of human neutrophils: a possible oxidase in the respiratory burst. Biochem J 194:599–606
Wood PM (1974) The redox potential of the system oxygensuperoxide. FEBS Lett 44:22–24
Royer-Pokora B, Kunkel LK, Monaco AP, Goff SC, Newburger PE, Baehner RL, Cole FS, Curnutte JT, Orkin SH (1986) Cloning the gene for an inherited human disorder — chronic granulomatous disease — on the basis of its chromosomal location. Nature 322:32–38
Dinauer MC, Orkin SH, Brown R, Jesaitis AJ, Parkos CA (1987) The glycoprotein encoded by the X-linked chronic granulomatous disease locus is a component of the neutrophil cytochromeb complex. Nature 327:717–720
Teahan C, Towe P, Parker P, Totty N, Segal AW (1987) The X-linked chronic granulomatous disease gene codes for the Β-chain of cytochromeb 245. Nature 327:720–721
Segal AW (1987) Absence of both cytochromeb-245 subunits from neutrophils in X-linked chronic granulomatous disease. Nature 326:88–91
Parkos CA, Allen RA, Cochrane CG, Jesaitis AJ (1987) Purified cytochromeb from human granulocyte plasma membrane is composed of two polypeptides with relative molecular weights of 91000 and 22000. J Clin Invest 80:732–742
Yamaguchi T, Hayakawa T, Kaneda M, Kakinuma K, Yoshikawa A (1989) Purification and some properties of the small subunit of cytochromeb 558 from human neutrophils. J Biol Chem 264:112–118
Nugent JH, Gratzer W, Segal AW (1989) Identification of the haem-binding subunit of cytochromeb 245. Biochem J 264:921–924
Nakamura M, Sendo S, Van Zwieten R, Koga T, Roos D, Kanegasaki S (1988) Immunocytochemical discovery of the 22-to 23-Kd subunit of cytochromeb 558 at the surface of human peripheral phagocytes. Blood 72:1550–1552
Kleinberg ME, Rotrosen D, Malech HL (1989) Asparaginelinked glycosylation of cytochromeb 558 large subunit varies in different human phagocytic cells. J Immunol 143:4152–4157
Imajoh-Ohmi S, Tokita K, Ochiai H, Nakamura M, Kanegasaki S (1992) Topology of cytochromeb 558 in neutrophil membrane analyzed by anti-peptide antibodies and proteolysis. J Biol Chem 267:180–184
Rotrosen D, Kleinberg ME, Nunoi H, Leto T, Gallin JI, Malech HL (1990) Evidence for a functional cytoplasmic domain of phagocyte oxidase cytochromeb 558. J Biol Chem 265:8745–8750
Quinn MT, Parkos CA, Walker L, Orkin SH, Dinauer MC (1989) Association of Ras-related protein with cytochromeb of human neutrophils. Nature 342:198–200
Quinn MT, Mullen ML, Jesaitis AJ, Linner JG (1992) Subcellular distribution of the Rap1A protein in human neutrophils: Colocalization and cotranslocation with cytochromeb 559. Blood 79:1563–1573
Bokoch GM, Quilliam LA, Bohl BP, Jesaitis AJ, Quinn MT (1991) Inhibition of Rap1A binding to cytochromeb 558 of NADPH oxidase by phosphorylation of Rap1A. Science 254:1794–1796
Trahey M, McCormick F (1987) A cytoplasmic protein stimulates normal N-ras p21 GTPase, but does not affect oncogenic mutants. Science 238:542–545
Lomax KJ, Leto TL, Nunoi H, Gallin JI, Malech HL (1989) Recombinant 47-kilodalton cytosol factor restores NADPH oxidase in chronic granulomatous disease. Science 245:409–412
Eklund EA, Marshall M, Gibbs JB, Crean CD, Gabig TG (1991) Resolution of a low-molecular-weight G protein in neutrophil cytosol required for NADPH oxidase activation and reconstitution by recombinant Krev-1 protein. J Biol Chem 266:13964–13970
Cagan RH, Karnovsky ML (1964) Enzymic basis of the respiratory stimulation during phagocytosis. Nature 204:255–257
Markert J, Glass GA, Babior BM (1985) Respiratory burst oxidase from human neutrophils: purification and some properties. Proc Natl Acad Sci USA 82:3144–3148
Kakinuma K, Kaneda M, Chiba T, Ohnishi T (1986) Electron spin resonance studies on a flavoprotein in neutrophil plasma membranes: redox potentials of the flavin and its participation in NADPH oxidase. J Biol Chem 261:9426–9432
Doussière J, Vignais PV (1985) Purification and properties of an O .2 −-generating oxidase from bovine polymorphonuclear neutrophils. Biochemistry 24:7231–7239
Kakinuma K, Fukuhara Y, Kaneda M (1987) The respiratory burst oxidase of neutrophils. Separation of an FAD enzyme and its characterization. J Biol Chem 262:12316–12322
Doussière J, Vignais PV (1988) Immunological properties of O .2 −-generating oxidase from bovine neutrophils. FEBS Lett 234:362–366
Yea CM, Cross AR, Jones OTG (1990) Purification and some properties of the 45-kDa diphenylene iodonium-binding flavoprotein of neutrophil NADPH oxidase. Biochem J 265:95–100
Parkinson JF, Gabig TG (1988) Phagocyte NADPH-oxidase. Studies with flavin analogues as active-site probes in Triton X-100-solubilized preparations. J Biol Chem 263:8859–8863
Bromberg T, Pick E (1985) Activation of NADPH-dependent superoxide production in a cell-free system by sodium dodecyl sulfate. J Biol Chem 260:13539–13545
Sumimoto H, Sakamoto N, Nozaki M, Sakaki Y, Takeshige K, Minakami S (1992) Cytochromeb 558, a component of the phagocyte NADPH oxidase, is a flavoprotein. Biochem Biophys Res Commun 186:1368–1375
Sha'ag D, Pick E (1988) Macrophage-derived superoxidegenerating NADPH oxidase in an amphiphile-activated, cell-free system; partial purification of the cytosolic component and evidence that it may contain the NADPH-binding site. Biochim Biophys Acta 952:213–219
Segal AW, West I, Wientjes F, Nugent JHA, Chavan AJ, Haley B, Rosen RC, Scrace G (1992) Cytochromeb 245 is a flavocytochrome containing FAD and the NADPH-binding site of the microbicidal oxidase of phagocytes. Biochem J 284:781–788
Knoller S, Shpungin S, Pick E (1991) The membrane-associated component of the amphiphile-activated, cytosol-dependent superoxide-forming NADPH oxidase of macrophages is identical to cytochromeb 558. J Biol Chem 266:2795–2804
Karplus PA, Daniels MJ, Herriott JR (1991) Atomic structure of ferredoxin-NADP+ reductase: prototype for a structurally novel flavoenzyme family. Science 251:60–66
Bredt DS, Hwang PM, Glatt CE, Lowenstein C, Reed RR, Snyder SH (1991) Cloned and expressed nitric oxide synthase structurally resembles cytochrome P-450 reductase. Nature 351:714–718
Rotrosen D, Yeung CL, Leto TL, Malech HL, Kwong CH (1992) Cytochromeb 558: the flavin-binding component of the phagocyte NADPH oxidase. Science 256:1459–1462
Umeki S (1990) Human neutrophil cytosolic activation factor of the NADPH oxidase: characterization of activation kinetics. J Biol Chem 265:5049–5054
Cox JA, Jeng AY, Sharkey NA, Blumberg PM, Tauber AI (1985) Activation of the human neutrophil nicotinamide adenine dinucleotide phosphate (NADPH)-oxidase by protein kinase C. J Clin Invest 76:1932–1938
Bromberg Y, Pick E (1984) Unsaturated fatty acids stimulate NADPH-dependent superoxide production by cell-free system derived from macrophages. Cell Immunol 88:213–221
Heyneman RA, Vercauteren RE (1984) Activation of an NADPH oxidase from horse polymorphonuclear leukocytes in a cell-free system. J Leukocyte Biol 36:751–759
Umeki S (1990) Activation of the NADPH oxidase in a cellfree system from human neutrophils stimulated by phorbol myristate acetate. Life Sci 46:1111–1118
Babior BM, Kuver R, Curnutte JT (1988) Kinetics of activation of the respiratory burst oxidase in a fully soluble system from human neutrophils. J Biol Chem 263:1713–1718
Nunoi H, Rotrosen D, Gallin JI, Malech HL (1988) Two forms of autosomal chronic granulomatous disease lack distinct neutrophil cytosol factors. Science 242:1298–1301
Volpp BD, Nauseef WM, Clark RA (1988) Two cytosolic neutrophil oxidase components absent in autosomal chronic granulomatous disease. Science 242:1295–1297
Caldwell SE, McCall CE, Hendricks CL, Leone PA, Bass DA, McPhail LC (1988) Coregulation of NADPH oxidase activation and phosphorylation of a 48-kD protein(s) by a cytosolic factor defective in autosomal recessive chronic granulomatous disease. J Clin Invest 81:1485–1496
Bolscher BGJM, Van Zwieten R, Kramer IJM, Weening RS, Verhoeven AJ, Roos D (1989) A phosphoprotein of Mr 47000, defective in autosomal chronic granulomatous disease copurifies with one of two soluble components required for NADPH:O2 oxidoreductase activity in human neutrophils. J Clin Invest 83:757–763
Hayakawa T, Suzuki K, Suzuki S, Andrews PC, Babior EM (1986) A possible role for protein phosphorylation in the activation of the respiratory burst in human neutrophils. Evidence from studies with cells from patients with chronic granulomatous disease. J Biol Chem 261:9109–9115
Uhlinger DJ, Burnham DN, Lambeth JD (1991) Nucleotide triphosphate requirements for superoxide generation and phosphorylation in a cell-free system from human neutrophils. Sodium dodecyl sulfate and diacylglycerol activate independently of protein kinase C. J Biol Chem 266:20990–20997
Heyworth PG, Segal AW (1986) Further evidence for the involvement of a phosphoprotein in the respiratory burst oxidase of human neutrophils. Biochem J 239:723–731
Kramer IJM, Verhoeven AJ, Van der Bend RL (1988) Purified protein kinase C phosphorylates a 47-kDa protein in control neutrophil cytoplasts but not in neutrophil cytoplasts from patients with the autosomal form of chronic granulomatous disease. J Biol Chem 263:2352–2357
Badwey JA, Robinson JM, Heyworth PG, Curnutte JT (1989) 1,2-Dioctanoyl-sn-glycerol can stimulate neutrophils by different mechanisms. Evidence for a pathway that does not involve phosphorylation of the 47-kDa protein. J Biol Chem 264:20676–20682
Sha'afi RI, Molski TFP, Gomez-Cambronero J, Huang C-K (1988) Dissociation of the 47-kilodalton protein phosphorylation from degranulation and superoxide production in neutrophils. J Leukocyte Biol 43:18–27
Rotrosen D, Leto TL (1990) Phosphorylation of neutrophil 47-kDa cytosolic oxidase factor. Translocation to membrane is associated with distinct phosphorylation events. J Biol Chem 265:19910–19915
Clark RA, Volpp BD, Leidal KG, Nauseef WM (1990) Two cytosolic components of the human neutrophil respiratory burst oxidase translocate to the plasma membrane during cell activation. J Clin Invest 85:714–721
Heyworth PG, Curnutte JT, Nauseef WM, Volpp BD, Pearson DW, Rosen H, Clark RA (1991) Neutrophil nicotinamide adenine dinucleotide phosphate oxidase assembly. Translocation of p47-phox and p67-phox requires interaction between p47-phox and cytochromeb 558. J Clin Invest 87:352–356
Tyagi SR, Neckelmann N, Uhlinger DJ, Burnham DN, Lambeth JD (1992) Cell-free translocation of recombinant p47-phox, a component of the neutrophil NADPH oxidase: effects of guanosine 5′-O-(3-thiotriphosphate), diacylglycerol, and an anionic amphiphile. Biochemistry 31:2765–2774
Okamura N, Babior BM, Mayo LA, Peveri P, Smith RM, Curnutte JT (1990) The p67-phox cytosolic peptide of the respiratory burst oxidase from human neutrophils. Functional aspects. J Clin Invest 85:1583–1587
Fujimoto S, Smith RM, Curnutte JT, Babior BM (1989) Evidence that activation of the respiratory burst oxidase in a cell-free system from human neutrophils is accomplished in part through an alteration of the oxidase related 67-kDa cytosolic protein. J Biol Chem 264:21629–21632
Teahan CG, Totty N, Casimir CM, Segal AW (1990) Purification of the 47-kDa phosphoprotein associated with the NADPH oxidase of human neutrophils. Biochem J 267:485–489
Nauseef WM, Volpp BD, Clark RA (1990) Immunochemical and electrophoretic analyses of phosphorylated native and recombinant neutrophil oxidase component p47-phox. Blood 76:2622–2629
Tanaka T, Imajoh-Ohmi S, Kanegasaki S, Ishimura I (1990) A 63-kilodalton cytosolic polypeptide involved in superoxide generation in porcine neutrophils. J Biol Chem 265:18717–18720
Pilloud-Dagher M-C, Vignais PV (1991) Purification and characterization of an oxidase activating factor of 63 kilodaltons from bovine neutrophils. Biochemistry 30:2753–2760
Volpp BD, Nauseef WM, Donelson JE, Moser DR, Clark RA (1989) Cloning of the cDNA and functional expression of the 47-kilodalton cytosolic component of human neutrophil respiratory burst oxidase. Proc Natl Acad Sci USA 86:7195–7199
Leto TL, Lomax KJ, Volpp BD, Nunoi H, Sechler JMG, Nauseef WM, Clark RA, Gallin JI, Malech HL (1990) Cloning of a 67-kD neutrophil oxidase factor with similarity to a noncatalytic region of p60c-src. Science 248:727–730
Okamura N, Curnutte JT, Roberts RL, Babior BM (1988) Relationship of protein phosphorylation to the activation of the respiratory burst in human neutrophils. Defects in the phosphorylation of a group of closely related 48-kDa proteins in two forms of chronic granulomatous disease. J Biol Chem 263:6777–6782
Hall A (1990) The cellular functions of small GTP-binding proteins. Science 249:635–640
Abo A, Pick E, Hall A, Totty N, Teahan CG, Segal AW (1991) Activation of the NADPH oxidase involves the small GTP-binding protein p21racl. Nature 353:668–670
Knaus UG, Heyworth PG, Kinsella BT, Curnutte JT, Bokoch GM (1992) Purification and characterization of Rac 2. A cytosolic GTP-binding protein that regulates human neutrophil NADPH oxidase. J Biol Chem 267:23575–23582
Koppenol WH (1989) Generation and thermodynamic properties of oxyradicals. In: CRC Critical reviews in membrane lipid oxidation, vol 1. CRC Press, Boca Raton, pp 1–13
Cross AR, Parkinson JF, Jones OTG (1985) Mechanism of the superoxide-producing oxidase of neutrophils. O2 is necessary for the fast reduction of cytochromeb-245 by NADPH. Biochem J 226:881–884
Iizuka T, Kanegasaki S, Makino R, Tanaka T, Ishimura Y (1985) Pyridine and imidazole reversibly inhibit the respiratory burst in porcine and human neutrophils: evidence for the involvement of cytochromeb 558 in the reaction. Biochem Biophys Res Commun 130:621–626
Wainio WW, Greenlees J (1960) Complexes of cytochromec oxidase with cyanide and carbon monoxide. Arch Biochem Biophys 90:18–21
Ishimura Y, Ullrich V, Peterson JA (1971) Oxygenated cytochrome P-450 and its possible role in enzymic hydroxylation. Biochem Biophys Res Commun 42:140–146
Iizuka T, Kanegasaki S, Makino R, Tanaka T, Ishimura Y (1985) Studies on neutrophilb-type cytochrome in situ by low-temperature absorption spectroscopy. J Biol Chem 260:12049–12053
Butler J, Koppenol WH, Margoliash E (1982) Kinetics and mechanism of the reduction of ferricytochromec by the superoxide anion. J Biol Chem 257:10747–10750
Porter TD, Kasper CB (1986) NADPH-cytochrome P-450 oxidoreductase: flavin mononucleotide and flavin adenine dinucleotide domains evolved from different flavoproteins. Biochemistry 25:1682–1687
Crawford NM, Smith M, Bellissimo D, Davis RW (1988) Sequence and nitrate regulation of theArabidopsis thaliana mRNA encoding nitrate reductase, a metalloflavoprotein with three functional domains. Proc Natl Acad Sci USA 85:5006–5010
Ostrowski J, Barber MJ, Rueger DC, Miller BE, Siegel LM, Kredich NM (1989) Characterization of the flavoprotein moieties of NADPH-sulfate reductase fromSalmonella typhimurium andEscherichia coli. Physicochemical and catalytic properties, amino acid sequence deduced from DNA sequence ofcysJ, and comparison with NADPH-cytochrome P-450 reductase. J Biol Chem 264:15796–15808
Quinn MT, Mullen ML, Jesaitis AJ (1992) Human neutrophil cytochromeb contains multiple hemes. Evidence for heme associated with both subunits. J Biol Chem 267:7303–7309
Clark RA, Malech HL, Gallin JI, Nunoi H, Volpp BD, Pearson D, Nauseef WM, Curnutte JT (1989) Genetic variants of chronic granulomatous disease: prevalence of deficiencies of two cytosolic components of the NADPH oxidase system. N Engl J Med 321:647–652
Umeki S (1991) Topics in chronic granulomatous disease. Pediatrics 88:183–185
Heyworth PG, Shrimpton CF, Segal AW (1989) Localization of the 47-kDa phosphoprotein involved in the respiratoryburst NADPH oxidase of phagocytic cells. Biochem J 260:243–248
Ohno Y, Buescher ES, Roberts R, Metcalf JA, Gallin JI (1986) Reevaluation of cytochromeb and flavin adenine dinucleotide in neutrophils from patients with chronic granulomatous disease, and description of a family with probable autosomal recessive inheritance of cytochromeb deficiency. Blood 67:1132–1138
Francke U, Hsieh C-L, Foellmer BE, Lomax KJ, Malech HL, Leto TL (1990) Genes for two autosomal recessive forms of chronic granulomatous disease assigned to 1q25 (NCF2) and 7q11.23 (NCF1). Am J Hum Genet 47:483–492
Kalomiris EL, Bourguignon LYW (1989) Lymphoma protein kinase C is associated with the transmembrane glycoprotein, gp85, and may function in gp85-ankyrin binding. J Biol Chem 264:8113–8119
Dinauer MC, Pierce EA, Bruns GA, Curnutte JT, Orkin SH (1990) Human neutrophil cytochromeb light chain (p22-phox). Gene structure, chromosomal location, and mutations in cytochrome-negative autosomal recessive chronic granulomatous disease. J Clin Invest 86:1729–1737
Parkos CA, Dinauer MC, Jesaitis AJ, Orkin SH, Curnutte JT (1989) Absence of both the 91-kD and 22-kD subunits of human neutrophil cytochromeb in two genetic forms of chronic granulomatous disease. Blood 73:1416–1420
Dinauer MC, Pierce EA, Erickson RW, Muhlebach TJ, Messner H, Orkin SH, Seger RA, Curnutte JT (1991) Point mutation in the cytoplasmic domain of the neutrophil p22-phox cytochromeb subunit is associated with a nonfunctional NADPH oxidase and chronic granulomatous disease. Proc Natl Acad Sci USA 88:11231–11235
Dinauer MC, Curnutte JT, Rosen H, Orkin SH (1989) A missense mutation in the neutrophil cytochromeb heavy chain in cytochrome-positive X-linked chronic granulomatous disease. J Clin Invest 84:2012–2016
The International Chronic Granulomatous Disease Cooperative Study Group (1991) A controlled trial of interferon gamma to prevent infection in chronic granulomatous disease. N Engl J Med 324:509–516
Berton G, Zeni L, Cassatella NA, Rossi F (1986) Gamma interferon is able to enhance the oxidative metabolism of human neutrophils. Biochem Biophys Res Commun 138:1276–1282
Ezekowitz RAB, Okin SH, Newburger PE (1987) Recombinant interferon gamma augments phagocyte superoxide production and X-chronic granulomatous disease gene expression in X-linked variant chronic granulomatous disease. J Clin Invest 80:1009–1016
Sechler JMG, Malech HL, White CJ, Gallin JI (1988) Recombinant human interferon-γ reconstitutes defective phagocyte function in patients with chronic granulomatous disease of childhood. Proc Natl Acad Sci USA 85:4874–4878
Ezekowitz RAB, Dinauer MC, Jaffe HS, Orkin SH, Newburger PE (1988) Partial correction of the phagocyte defect in patients with X-linked chronic granulomatous disease by subcutaneous interferon gamma. N Engl J Med 319:146–151
Woodman RC, Erickson RW, Rae J, Jaffe HS, Curnutte JT (1992) Prolonged recombinant interferon-γ therapy in chronic granulomatous disease: evidence against enhanced neutrophil oxidase activity. Blood 79:1558–1562
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Umeki, S. Mechanisms for the activation/electron transfer of neutrophil NADPH-oxidase complex and molecular pathology of chronic granulomatous disease. Ann Hematol 68, 267–277 (1994). https://doi.org/10.1007/BF01695032
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01695032