1. Protein delivery by Pseudomonas type III secretion system: Ex vivo complementation of p67(phox)-deficient chronic granulomatous disease.
- Author
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Polack B, Vergnaud S, Paclet MH, Lamotte D, Toussaint B, and Morel F
- Subjects
- B-Lymphocytes enzymology, B-Lymphocytes microbiology, B-Lymphocytes pathology, Bacterial Proteins genetics, Bacterial Proteins metabolism, Biological Transport, Cell Line, Transformed, Cytosol enzymology, Cytosol metabolism, Granulomatous Disease, Chronic genetics, Granulomatous Disease, Chronic metabolism, Herpesvirus 4, Human genetics, Herpesvirus 4, Human physiology, Histidine Kinase, Humans, NADPH Oxidases metabolism, Phosphoproteins genetics, Phosphoproteins metabolism, Protein Kinases genetics, Recombinant Fusion Proteins metabolism, Virulence genetics, B-Lymphocytes metabolism, Genetic Complementation Test, Granulomatous Disease, Chronic enzymology, Phosphoproteins deficiency, Protein Kinases metabolism, Pseudomonas aeruginosa enzymology, Pseudomonas aeruginosa genetics, Pseudomonas aeruginosa metabolism
- Abstract
Bacterial type III secretion system drives the translocation of virulence factors into the cystosol of host target cells. In phagocytes and in Epstein-Barr virus immortalized B lymphocytes, NADPH oxidase generates O(-2) through an electron transfer chain the activity of which depends on the assembly of three, p67(phox), p47(phox) and p40(phox) cytosolic activating factors with Rac 1/2 and a membrane redox component, cytochrome b(558). In p67(phox) deficient chronic granulomatous disease (CGD) patients, p67-phox is missing and NADPH oxidase activity is abolished. ExoS is a virulence factor of Pseudomonas aeruginosa which is secreted via the type III secretion system: it was fused with p67(phox). Pseudomonas aeruginosa synthesized and translocated the hybrid ExoS-p67(phox) fusion protein into the cytosol of B lymphocytes via the type III secretion system. Purified ExoS-p67(phox) hybrid protein was as efficient as normal recombinant p67(phox) in cell-free reconstitution of NADPH oxidase activity. Therefore, ExoS-p67(phox) was transferred via the type III secretion system of Pseudomonas aeruginosa into the cytosol of B lymphocytes from a p67(phox)-deficient CGD patient and functionally reconstituted NADPH oxidase activity. In the complementation process, ExoS acted as a molecular courier for protein delivery: the reconstitution of an active NADPH oxidase complex suggests type III secretion system to be a new approach for cellular therapy., (Copyright 2000 Academic Press.)
- Published
- 2000
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