Paul A. Johnston
Research Associate Professor, Project Manager
3501 5th Ave
Pittsburgh, PA 15260
|B.S. University of East Anglia, Norwich, England
Ph.D. Biochemistry, University of East Anglia, Norwich, England, 1983
1983-1984. North Carolina University, Chapel Hill, NC. Department of Pharmacology, School of Medicine. Dr. Gene A. Scarborough. Biochemistry.
1984-1987. Duke University Medical Center, Durham, NC. Department of Pathology. Drs. Dolph O. Adams and Tom Hamilton. Immunology.
1988-1990. University of Texas Southwestern, Howard Hughes Medical Institute, Dallas, TX. Dr. Thomas Sudhof. Molecular Biology and Biochemistry.
|Macrophage activation, Drug discovery, Assay development, High throughput screening, High content screening, Cell based screening, Image based assay development and implementation.
|1. Johnston, P.A. and A. Coddington (1982). Multiple Drug Resistance in the Fission Yeast Schizosaccharomyces pombe: Evidence for the Existence of Pleiotropic Mutations Affecting Energy Dependent Transport systems. Mol. Gen. Genet. 185: 311-314.
2. Johnston, P.A. and A. Coddington (1983). Multiple Drug Resistance in the Fission Yeast Schizosaccharomyces pombe: Correlation between Drug and Amino Acid Uptake and Membrane ATPase Activities. Current Genetics 7: 299-307.
3. Johnston, P.A. and A. Coddington (1984). Drug Resistance in the Fission Yeast Schizosaccharomyces pombe: Pleiotropic Mutations Affecting the Oleic Acid and Sterol Composition of Cell Membranes. Current Genetics 8: 37-43.
4. Johnston, P.A., D.O. Adams, and T.A. Hamilton (1984). Fc-receptor-Mediated Protein Phosphorylation in Murine Peritoneal Macrophages. Biochem. Biophys. Res. Commun. 124: 197-202.
5. Johnston, P.A., D.O. Adams, and T.A. Hamilton (1985). Regulation of the Fc-receptor-mediated Respiratory Burst: Treatment of Primed Murine Peritoneal macrophages with Lipopolysaccharide Selectively Inhibits H202 Secretion Stimulated by Immune Complexes. J. Immunol.135: 513-518.
6. Johnston, P.A., D.O. Adams and T.A. Hamilton (1986). Regulation of Respiratory Burst in Murine Peritoneal macrophages: Differential Sensitivity to Phorbol Diesters by Macrophages in Different Stages of Functional Activation. Cell. Immunol. 100: 400-410.
7. Johnston, P.A., M.M Jansen, S.D. Somers, D.O. Adams and T.A. Hamilton (1987). Ligands for the Scavenger Receptor on Murine Peritoneal Macrophages Induce Expression of a Set of Early Proteins. J. Immunol. 138: 1551-1558.
8. Johnston, P.A. S.D. Somers and T.A. Hamilton (1987). Expression of a 120 Kilodalton Protein During Tumoricidal Activation in Murine Peritoneal Macrophages. J. Immunol. 138: 2739-2744.
9. Introna, M., R.C. Bast, Jr., P.A. Johnston, D.O. Adams and T.A. Hamilton (1987). Homologous and Heterologous Desensitization of Proto-Oncogene cFos Expression in Murine Peritoneal Macrophages. J. Cell. Physiol. 131: 36-42.
10. Johnston, P.A., T.J. Koerner, M.M. Jansen and T.A. Hamilton (1989). Expression of Macrophage p120 Depends upon Early Protein Synthesis. J. Immunol. 142: 2728-2735.
11. Johnston, P.A., G.A. Reynolds, S. Wasserman and T.C. Sudhof (1990). Two Novel Annexins from Drosophila Melanogaster. Cloning, Characterization and Differential Expression in Development. J. Biol. Chem 265: 11382-11388.
12. Johnston, P.A., F. Yu, G.A. Reynolds, H.L. Yin, C.R. Moomaw, C.A. Slaughter C.A. and T.A. Sudhof (1990). Purification and Expression of gCap39. An Intracellular and Secreted Ca2+-Dependent Actin-Binding Protein Enriched in Mononuclear Phagocytes. J. Biol. Chem. 265: 17946-17952.
13. Yu, Fu-Xin, P.A. Johnston, T.C. Sudhof and H.L. Yin (1990). gCap39, a Calcium Ion and Polyphosphoinositide-Regulated Actin Capping Protein. Science 250: 1413-1415.
14. Johnston, P.A. and T.C. Sudhof (1990). Evolutionary Conservation and Structure-Function Relationships in the Calelectrins (Annnexins). Biochem. Soc. Trans. 18: 1097-1098.
15. Perrin, M., P.A. Johnston, T. Ozcelik, R. Jahn, U. Franke and T.C. Sudhof (1991). Structural and Functional Conservation of Synaptotagnin p65) in Drosophila and Humans. J. Biol. Chem. 266: 615-622.
16. Mignery, G.A., P.A. Johnston and T.C. Sudhof (1992). Mechanism of Ca2+ Inhibition of Inositol 1,4,5-Trisphosphate (InsP3) Binding to the Cerebellar InsP3 Receptor. J. Biol. Chem. 267: 7450-7455.
17. Daley, M.J., G. Furda, R. Dougherty, P.A. Coyle, T.J. Williams and P.A. Johnston (1992). Potentiation of Antibiotic Therapy for Bovine Mastitis by Recombinant Bovine Interleukin-2. J. Dairy Science 75: 3330-3338.
18. Tao, W.T., R. Dougherty, P.A. Johnston and W. Pickett (1993). Recombinant Bovine GM-CSF Primes Platlet Activating Factor, rHuIL-8 but not rBoIL-1 Induced Bovine Neutrophil Degranulation. J. Leukocyte Biology 53: 679-684.
19. Bolnet, C., P.A. Johnston, A. Kemper, C. Ricks, & J. Petite (1995). Influence of Avian Con A Induced Splenocyte Conditioned Media on Cells of the Hematopoietic Lineage. J. Poultry Sci. 74: 1102-1116.
20. Johnston, P. A. and P. A. Johnston (2002). Cellular Platforms for HTS: three case studies. Drug Discovery Today 7: 353-363.
21. Pratt S, Shepard RL, Kandasamy RA, Johnston PA, Perry W 3rd, and Dantzig AH. (2005). The multidrug resistance protein 5 (ABCC5) confers resistance to 5-fluorouracil and transports its monophosphorylated metabolites. Mol. Cancer Ther. 4: 855-63.
22. Razonable RR, Hennault M, Lee LN, Laethem C, Johnston PA, Watson HL and Paya CV. (2005). Secretion of Proinflammatory Cytokines and Chemokines during Amphotericin B Exposure is Mediated by Coactivation of Toll-Like Receptors 1 and 2. Antimicrobial Agents and Chemotherapy. 49: 1617-1621
23. A. H. Gough and P. A. Johnston. (2006) Requirements, Features and Performance of High Content Screening Platforms, in High Content Screening: A Powerful Approach to Systems Cell Biology and Drug Discovery. Methods in Molecular Biology 356: 41-61. Humana Press, Totowa, NJ. Editors: D. L. Taylor, J. R. Haskins and K.A. Giuliano
24. R.G.Williams, R.Kandasamy, D.Nickischer, O.J. Trask, C.Laethem, P.A Johnston, and P.A. Johnston. (2006). “Generation and Characterization of a Stable MK2-EGFP Cell Line and Subsequent Development of a High Content Imaging Assay on the Cellomics ArrayScan® Platform to Screen for p38 MAPK Inhibitors” in Methods in Enzymology 414: 364-89. “Automated Microscopy Screening” Elsevier/Academic Press Edited by Jim Inglese.
25. O.J. Trask, A.Baker, R.G.Williams, D.Nickischer, R. Kandasamy, C. Laethem, P. A. Johnston, and P. A. Johnston. (2006). “Assay Development and Case History of a 32K Biased Library High Content MK2-EGFP Translocation Screen to Identify p38 MAPK Inhibitors on the ArrayScan® 3.1 Imaging Platform” in Methods in Enzymology 414: 419-39. “Automated Microscopy Screening” Elsevier/Academic Press Edited by Jim Inglese.
26. D. Nickischer, C. L. Laethem, O. J. Trask, Jr., R. G. Williams, R. Kandasamy, P. A. Johnston, and P. A. Johnston. (2006). “Development and implementation of three MAPK signaling pathway imaging assays to provide MAPK module selectivity profiling for kinase inhibitors; MK2-EGFP translocation, c-Jun & ERK activation” in Methods in Enzymology 414: 389-418. “Automated Microscopy Screening” Elsevier/Academic Press Edited by Jim Inglese.
27. Arnold DM, Foster C, Huryn DM, Lazo JS, Johnston PA, Wipf P. (2007) Synthesis and Biological Activity of a Focused Library of Mitogen-activated Protein Kinase Phosphatase Inhibitors. Chem. Biol. Drug Des. 69:23-30.
28. Brisson-Tierno M*, Johnston PA*, Foster C, Skoko JJ, Shun TY, Lazo JS. (2007) Development and Optimization of high-throughput in vitro protein phosphatase screening assays for drug discovery. Nature Protocols. Manuscript in press. * Authors contributed equally to this work.
29. Johnston PA, Foster CA, Shun TY, Skoko JJ, Shinde S, Wipf P, and Lazo JS. (2007) MKP-1 Dual Specificity Protein Phosphatase Inhibitor Probe Assessment: Secondary Assay Follow Up of Actives Identified in an NIH Molecular Library Screening Center Network (MLSCN) MKP-1 HTS. Manuscript revised and re-submitted.
30. Paul A. Johnston, Jennifer Phillips, Tong Ying Shun, Sunita Shinde, John S. Lazo, Donna, M. Huryn, Michael M. Myers, Boris I. Ratnikov, Jeffrey W. Smith, Ying Su, RusselDahl, Nicholas D. P. Cosford, Sergey A. Shiryaev and Alex Y. Strongin, “High throughput screening (HTS) identifies novel and specific uncompetitive inhibitors of the two-component NS2B-NS3 proteinase of West Nile virus”, Assay Drug Dev Technologies. 5: p 737-750
|Ongoing Research Support
NIH 1U54MH074411-01 (Lazo) 2005-2007 $34,800 1.8 calendar
Role: Manager of the University of Pittsburgh Molecular Library Screening Center (PMLSC) 15%
Leader of Cores 1 & 2, the assay development, implementation and high throughput screening cores.
Member of Core 5, the PMLSC administrative core.
The goals of this application are to develop, implement, and optimize robust optical methods for the detection, characterization, and refinement of small molecules with desired biological and pharmacological properties in cell-free, cell-based, and model organism-based assays as assigned by the MLSCN steering group.
Pending Research Support
NIH (Pak) $23,200 1.2 calendar
Role: Co-Investigator (10%)
Identification of novel therapeutic agents for Alpha-1-antitrypsin deficiency using C. elegans
NIH (Silverman) 12/01/07 – 11/30/12 $23,215 1.2 calendar
Role: Co-Investigator (10%)
High Throughput Genetic and Drug Screens for Alpha-1-Antitrypsin Deficiency
NIH/NINDS R21 NS059369-01 (Amara) 03/01/07 – 02/29/08 $52,634 1.2 calendar
Fluorescent pH Biosensor HTS Assays for Modulators of CNS Glutamate Transporters
The main goal of this proposal is to identify new classes of neuroprotective drugs that will lead to better treatments for seizures and stroke, and to prevent the neuronal death that is the hallmark of diseases such as Alzheimer’s disease and amyotrophic lateral sclerosis (ALS).
NIH R03 PAR-06-545 (Johnston) 10/01/04 – 09/30/08 $25,000 0.6 calendar
High Throughput Screening Assay to Measure the Generation of the Reactive Oxygen Intermediate Hydrogen Peroxide through Redox Cycling of Compounds in the presence of Dithiothreitol (DDT): Profiling the MLSCN Small Molecule Library to Identify Redox-based Promiscuous Inhibitors of Enzymes with Active Site Cysteines e.g. Phosphatases and Proteases
A simple homogeneous 384-well format biochemical HTS assay to profile the generation of H2O2 by compounds from the NIH small molecule library incubated with reducing agents has been developed and implemented. We will annotate the PubChem database with this data, and a simple cross target query of the PubChem database will alert investigators that these compounds exhibit non-selective and/or promiscuous bioactivity profiles and are false positives arising from their assay format that have little or no potential as probes. Hit compounds from a phenotypic cell based assay that are also active the H2O2 generation assay, might warrant further investigation of redox signaling events as a potential contributor to the altered phenotype.
|Obtained both a B.Sc. with Honors (2.1) (1978) and a Ph.D. (1983) in Biochemistry, from the University of East Anglia in Norwich, England. Subsequent postdoctoral positions in the department of Pharmacology at the University of North Carolina, the Pathology department of Duke University, and at the Howard Hughes Institute of the University of Texas Southwestern have provided a diversity of experience in biochemistry, molecular biology, cell biology, immunology, protein purification and recombinant protein expression.
I am a resourceful and productive scientific manager with fourteen years of drug discovery experience in the Pharmaceutical and Biotechnology industry. I was a Senior Research Immunologist at American Cyanamid, a Group Leader & Manager of the Embryology group at Embrex Inc., and Senior Project/Screen Leader & Research Advisor at Sphinx Pharmaceuticals/Eli Lilly Research Triangle Park. In the summer of 2005, I joined the University of Pittsburgh School of Medicine Department of Pharmacology to set up and manage a drug discovery center, one of the recipients of the NIH Roadmap Initiative Molecular Library Screening Center Network (MLSCN) grants.
An innovator of cell based approaches to lead generation and optimization, and recognized leader in the development and implementation of high content imaging technology to drug discovery. I have developed and run an in vitro ADME/Tox hit characterization program. I managed a group dedicated to lead generation; developing assays, running screens, implementing high content imaging assays, performing new technology evaluations, and providing cell culture support for all cell-based HTS.