Professor Emeritus, Department of Clinical Microbiology and Immunology, Sackler Faculty of Medicine, Tel Aviv University
Julius Friedrich Cohnheim Laboratory of Phagocyte Research
Member, Editorial Board, The FASEB Journal
Member, Editorial Board, International Journal of Hematology Research
Education
| 1961-1965 | M.D. | Hebrew University Hadassah Medical School |
| 1965-1967 | Research Fellow in Experimental Pathology | C Scripps Research Foundation, La Jolla, California, U.S.A. |
| 1967-1970 | Ph.D. in Immunology | University of London, London, U.K. |
| 1980-1981 | Research Fellow in Immunology | National Institutes of Health, Bethesda, MD |
Assembly of the Superoxide-Generating NADPH Oxidase Complex in Health and Disease
We are studying the production of reactive oxygen species (ROS) by phagocytes. ROS are generated by an enzyme complex, known as the NADPH oxidase. Our group is responsible for many of the seminal advances in the biochemistry and molecular biology of the NADPH oxidase complex, including: the standard micro-assay for the measurement of ROS (991 citations); the development of the first cell-free system of ROS production; the discovery of the cytosolic oxidase components (673 citations); the discovery of the role of the small GTPase Rac in oxidase activation (832 citations); the introduction of "peptide walking" to identify sites of protein-protein interaction, and the construction of chimeric cytosolic oxidase activators. The laboratory is equipped for the performance of advanced biochemical and molecular biology techniques.
The most recent interest of our group is focused on the mapping of the hotspots of interaction between the catalytic oxidase component Nox2 and the cytosolic activator p67phox. We found that the dehydrogenase region of Nox2 (residues 288-570) contains a Cys-Gly-Cys (CGC) triad (residues 369-371), which serves as a binding site for p67phox. This finding is based on a novel methodology, designed by us, in which we measure the binding of recombinant p67phox to an array of synthetic overlapping peptides covering the sequence of the dehydrogenase region of Nox2. Two Nox2 peptides that share the CGC triad, at their C- and N-termini, respectively, were found to bind p67phox. “Mutating” either C369 or C371 to R resulted in loss of p67phox binding. Chemical reduction of CGC-containing peptides also led to loss of binding. Linking the two cysteines by a disulfide bond resulted in a marked increase in binding. We concluded that binding of p67phox to the catalytic component of the NADPH oxidase complex is redox regulated and involves the establishment of disulfide bonds between p67phox and Nox2. The CGC triad might have a dual role by acting both as a protein disulfide isomerase (PDI) and by providing the cysteines for the establishment of disulfide bonds with p67phox. This novel hypothesis rests on the evidence that the CGC motif mimics functionally and structurally the CGHC catalytic site of members of the PDI family. Recently, we showed that a recombinant Nox2 construct possesses.
PDI activity, exhibits limited sequence similarity with PDIA3, and reacts with an anti-PDIA3 antibody.These findings have a key in vivo equivalent because a C369R mutation in human Nox2 causes Chronic Granulomatous Disease (CGD), an inborn defect resulting in the inability of phagocytes to produce ROS, leading to the failure to resist infections by bacteria and fungi.
Publications
Pick, E. When charge is in charge - "Millikan" for leukocyte biologists. J. Leukoc. Biol. 87, 537-540, 2010
Mizrahi, A., Berdichevsky, Y., Casey, P. J., and Pick, E. A prenylated p47phox-p67phox-Rac1 chimera is a quintessential NADPH oxidase activator. Membrane association and functional capacity. J. Biol. Chem. 285, 25485-25499, 2010
Hayee, B., Antonopoulos, A., Murphy, E. J., Rahman, F. Z., Sewell, G., Smith, B. N., McCartney, S., Furman, M., Hall, G., Bloom, S. L., Haslam, S. M., Morris, H. R., Boztug, K., Klein, C., Winchester, B., Pick, E., Linch, D. C., Gale, R. E., Smith, A. M., Dell, A., and Segal, A. W. G6PC3 mutations are associated with a major defect of glycosylation: A novel mechanism for neutrophil dysfunction. Glycobiology 21, 914-924, 2011
Pick, E., and Dahan, I. Strategies for identifying synthetic peptides to act as inhibitors of NADPH oxidases, or "All that you did and did not want to know about Nox inhibitory peptides". Cell. Mol. Life Sci., 69:2283-305, 2011
Bosco, E., Marchioni, F., Kumar, S., Biesiada, J., Kordos, M., Szczur, K., Meller, J., Seibel, W., Mizrahi, A., Pick, E., Filippi, M-D., and Zheng, Y. Rational design of small molecule inhibitors targeting the Rac GTPase - p67phox signaling axis in inflammation. Chem. Biol., 19:228-242, 2012
Dahan, I., Molshanski-Mor, S., and Pick, E. Inhibition of NADPH oxidase activation by peptides mapping within the dehydrogenase region of Nox2-A “peptide walking” study. J. Leukoc. Biol., 91:501-515, 2012
Dahan, I., and Pick, E. Strategies for identifying synthetic peptides to act as inhibitors of NADPH oxidases, or "All that you did and did not want to know about Nox inhibitory peptides". Cell. Mol. Life Sci. 69, 2283-2305, 2012
Roos, D., van Buul, J. D., Tool, A. T. J., Matute, J. D., Marchal, C. M., Hayee, B., Köker, M. Y., de Boer, M., van Leeuwen, K., Segal, A. W., Pick, E., and Dinauer M. C. Two CGD families with a hypomorphic mutation in the activation domain of p67-phox. J. Clin. and Cell. Immunol. in press, 2014
Chapter/Review
Pick, E., Cell-Free NADPH Oxidase Activation Assays - "In Vitro Veritas", In Neutrophil Methods and Protocols, 2nd Edition (Quinn, M. T., and DeLeo, F. R., eds), 339-403. Humana Press, 2014
Pick, E., Role of the Rho GTPase Rac in the activation of the phagocyte NADPH oxidase: Outsourcing a key task. Small GTPases, 5(1), 2014
Bechor, E., Dahan, I., Fradin, T., Berdichevsky, Y., Zahavi, A., Federman Gross, A., Rafalowski, M., and Pick, E. The dehydrogenase region of the NADPH oxidase component Nox2 acts as a protein disulfide isomerase (PDI) resembling PDIA3 with a role in the binding of the activator protein p67phox. Front. Chem. 3:3, 2015
Dahan, I., Smith S. M. E., and Pick, E. A Cys-Gly-Cys triad in the dehydrogenase region of Nox2 plays a key role in the interaction with p67phox. J. Leukoc. Biol. 98: 859-874, 2015
Pick, E. Absolute and relative activity values in assessing the effect of NADPH oxidase inhibitors. Antiox. Redox Signal. 23: 1250-1251, 2015
