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- Title
MOLECULAR APPROACHES TO THE STUDY OF THE PROTEIN OF 67 KDA (P67PHOX) OF NADPH OXIDASE SYSTEM OF THE PHAGOCYTIC CELLS.
- Authors
Arias, Andrés A.; Dinauer, Mary C.; Jiabin Ding; Rugeles, María T.; Patiño, Pablo J.
- Abstract
The NADPH oxidase system of the phagocytes cells has a central role for their microbicidal activity. The activation of this system is started by the translocation of cytosolic proteins p67phox, p47phox and p40phox towards the membrane to be in contact with flavocytochrome b558, inducing the superoxide anion generation. The present work presents a molecular study of NADPH oxidase system based on the findings of mutations in the p67phox gene of patients with chronic granulomatous disease (CGD) and some possible polymorphisms found in healthy humans. Mutations were generated in the p67phox cDNA by site-directed mutagenesis and then expressed in a COS-phox cells. The data collected in this work suggest that Lys19, Lys20 and Asp21 of the p67phox Nterminal are essential for the activation of the oxidase system while the C-terminal of this protein (residues 398-526) is not necessary for the O2- generation using this transgenic system. On the other hand, the changes Val166Ile, Pro329Ser and His389Gln corresponding to possible polymorphisms of normal individuals did not produce alterations in the of p67phox function, therefore these do not imply any genetic risk to produce deficiencies in the activation of NADPH oxidase system. In conclusion, the cellular model COS-phox represents a new system of complete cells, easily transfectable that serves to study the NADPH oxidase system function. Finally, the findings with the mutants of p67phox allow us to advance in the knowledge on the molecular mechanisms involved in the interaction of the cytosolic components of this system.
- Subjects
PHAGOCYTES; PROTEINS; SUPEROXIDES; GENETIC mutation; CHRONIC granulomatous disease; GENETIC polymorphisms; MUTAGENESIS
- Publication
Revista VacciMonitor (Vacunología y Temas Afines), 2002, Vol 11, Issue 4, p1
- ISSN
1025-028X
- Publication type
Article