Abstract

The respiratory burst oxidase is the enzyme responsible for the production of O2- by stimulated phagocytes. A highly complex enzyme composed of many component parts, it catalyzes the oneelectron reduction of oxygen to O 2- at the expense of NADPH. One of its components is an NADPH-binding polypeptide that is found in the cytosol of resting neutrophils but is transferred to the plasma membrane when the cells are activated, thereby assembling the electron transport chain of the enzyme. The goals of the project described in this application are the purification and characterization of this NADPH-binding component and the study of its interaction with other components of the oxidase. Through this work it may ultimately be possible to develop a sufficiently detailed molecular understanding of the operation of the respiratory burst oxidase to allow the development of a new class of anti-inflammatory agents that work by specifically controlling the production of reactive oxidants by phagocytes.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI030742-03
Application #
3145787
Study Section
Hematology Subcommittee 2 (HEM)
Project Start
1991-01-01
Project End
1995-12-31
Budget Start
1993-01-01
Budget End
1993-12-31
Support Year
3
Fiscal Year
1993
Total Cost
Indirect Cost

  Institution

Name
Scripps Research Institute
Department
Type
DUNS #
City
La Jolla
State
CA
Country
United States
Zip Code
92037

  Publications

Smith, R M; Connor, J A; Chen, L M et al. (1996) The cytosolic subunit p67phox contains an NADPH-binding site that participates in catalysis by the leukocyte NADPH oxidase. J Clin Invest 98:977-83
Gottlieb, R A; Giesing, H A; Engler, R L et al. (1995) The acid deoxyribonuclease of neutrophils: a possible participant in apoptosis-associated genome destruction. Blood 86:2414-8
Gottlieb, R A; Giesing, H A; Zhu, J Y et al. (1995) Cell acidification in apoptosis: granulocyte colony-stimulating factor delays programmed cell death in neutrophils by up-regulating the vacuolar H(+)-ATPase. Proc Natl Acad Sci U S A 92:5965-8
Park, J W; Benna, J E; Scott, K E et al. (1994) Isolation of a complex of respiratory burst oxidase components from resting neutrophil cytosol. Biochemistry 33:2907-11
el Benna, J; Ruedi, J M; Babior, B M (1994) Cytosolic guanine nucleotide-binding protein Rac2 operates in vivo as a component of the neutrophil respiratory burst oxidase. Transfer of Rac2 and the cytosolic oxidase components p47phox and p67phox to the submembranous actin cytoskeleton during oxidase J Biol Chem 269:6729-34
el Benna, J; Faust, L P; Babior, B M (1994) The phosphorylation of the respiratory burst oxidase component p47phox during neutrophil activation. Phosphorylation of sites recognized by protein kinase C and by proline-directed kinases. J Biol Chem 269:23431-6
Park, J W; Babior, B M (1993) Effects of diacylglycerol on the activation and kinetics of the respiratory burst oxidase in a cell-free system from human neutrophils: evidence that diacylglycerol may regulate nucleotide uptake by a GTP-binding protein. Arch Biochem Biophys 306:119-24
Foroozan, R; Ruedi, J M; Babior, B M (1992) The reduction of cytochrome b558 and the activity of the respiratory burst oxidase from human neutrophils. J Biol Chem 267:24400-7
Babior, B M (1992) The respiratory burst oxidase. Adv Enzymol Relat Areas Mol Biol 65:49-95
Umei, T; Babior, B M; Curnutte, J T et al. (1991) Identification of the NADPH-binding subunit of the respiratory burst oxidase. J Biol Chem 266:6019-22