Jeffrey Brodsky

Professor, Avinoff Chair of Biological Sciences

Protein quality control

Jeffrey Brodsky
Office: (412) 624-4831
Lab: (412) 624-4830
A321 Langley Hall
4249 Fifth Avenue
Pittsburgh, PA 15260
Education

Dr. Brodsky received his Ph.D. in 1990 with Guido Guidotti at Harvard University, performed his postdoctoral studies with Randy Schekman at the University of California, Berkeley, and joined the Department in 1994.

Figure 1.  A specific protein in yeast can be visualized using fluorescence microscopyFigure 1. A specific protein in yeast can be visualized using fluorescence microscopy

All secreted proteins, and most that ultimately reside within the cell, must traverse the secretory pathway, a network of intracellular organelles housing the “machines” that help secreted proteins mature.Critical components of these machines are a class of proteins known as molecular chaperones, some of which are associated with the endoplasmic reticulum (ER). If, however, protein folding is inefficient or slow, a secreted protein may be targeted for destruction by a process we termed ER Associated Degradation, or ERAD. During ERAD, proteins are selected as being defective, are modified with ubiquitin, and are degraded by the proteasome, a multi-catalytic protease that resides in the cytoplasm. Molecular chaperones are required for ERAD by “deciding” whether a protein is sufficiently mature to transit through the secretory pathway. Molecular chaperones can also direct ERAD substrates to the proteasome. The importance ofunderstanding the molecular mechanism of ERAD and molecular chaperone action is underscored by the fact that several human diseases—including cystic fibrosis, heart and liver disease, diabetes, and neurodegenerative diseases—can arise from defects in chaperone-mediated folding of secreted proteins and/or the ERAD pathway.

Figure 2.  A transcriptional profile analysis of yeast expressing CFTR, the protein that when mutated gives rise to cystic fibrosisFigure 2. A transcriptional profile analysis of yeast expressing CFTR, the protein that when mutated gives rise to cystic fibrosisFor our studies, the Brodsky laboratory primarily utilizes a model eukaryotic organism, the yeast Saccharomyces cerevisiae (Figure 1). Yeast possess the same intracellular membrane organization and molecular chaperones as human cells but are amenable to rapid genetic analysis. Moreover, the basic machinery required for ERAD is completely conserved between yeast and humans. Current research in the Brodsky laboratory is directed toward understanding how molecular chaperones in the ER and the cytoplasm facilitate ERAD and protein folding in the cell. Human proteins expressed heterologously in yeast, such as CFTR (Figure 2), are being examined as substrates for ERAD and chaperone-mediated folding. Data derived from our genetic studies are complemented by biochemical assays that recapitulate specific steps in the ERAD pathway, and by studies using cells from higher organisms in which disease phenotypes are more relevant. In parallel, we have identified and characterized small molecule modulators of specific molecular chaperones, some of which have potent inhibitory effects on cancer cells and on the replication of human viruses. New Roman";"> 

Students may find the Metabolic Pathways and Regulation (BIOSC1820) Course Website useful.

Recent Publications
  • Buck, T.M and J.L. Brodsky.  Escaping the endoplasmic reticulum: Why does a molecular chaperone leave home for greener pastures?  EMBO J. (2015), In press

  • Zacchi, L.F., J.J. Caramelo, A.A. McCracken, and J.L. Brodsky.  Endoplasmic Reticulum Associated Degradation and Protein Quality Control.  Encyclopedia of Cell Biology (R.A. Bradshaw and P. Stahl, Eds.), Elsevier.  (2015) In press

  • Wipf, P., Eyer, B.R., Yamaguchi, Y., Zhang, F., Neal, M.D., Sodhi, C.P., Good, M., Branca, M., Prindle, T., Jr., Lu, P., Brodsky, J. L., and Hackam, D. J. Synthesis of anti-Inflammatory α- and β-Linked Acetamidopyranosides as Inhibitors of Toll-Like Receptor 4 (TLR4). Tetrahedron Lett. (2015), In press

  • Li, H., H.C. Wu, Z. Liu, L.F. Zacchi, J.L. Brodsky, and M. Zolkiewski. Intracellular Complexes of the Early-Onset Torsion Dystonia-Associated AAA+ ATPase TorsinA. SpringerPlus (2014) 3:743

  • Howe, M.K., K. Bodoor, D.A. Carlson, P.F. Hughes, D.R. Loiselle, A.M. Jaeger, D.B. Darr, J.L. Jordan, L.M. Hunter, E.T. Molzberger, T.A. Gobillot, D.J. Thiele, J.L. Brodsky, N. Spector and T.A.J. Haystead.  Identification of a Novel Allosteric Small Molecule Inhibitor of the Inducible Form of Heat Shock Protein 70.  Chemistry & Biology (2014) 21: 1648–1659

  • Brodsky, J.L. and P.L. Clark.  Protein Folding in the Cell, From Atom to Organism.  FASEB J. (2014) 28: 5034-5038

  • Ireland, A.W., T.A. Gobillot, T. Gupta, S.P. Seguin, M. Liang, L. Resnick, K. Lloyd, M.T. Goldberg, A. Manos-Turvey, J.M. Pipas, P. Wipf, and J.L. Brodsky.  Synthesis and Structure Activity Relationships of Small Molecule Inhibitors of the Simian Virus 40 T Antigen Oncoprotein, an Anti-Polyomaviral Target.  Bioorg. Med. Chem. (2014) 22: 6490-6502

  • Surlow, B.A., B. Cooley, P.G. Needham, J.L. Brodsky, and J. Patton-Vogt.  Loss of Ypk1 accelerates Plb1-mediated phosphatidylcholine deacylation in S. cerevisiae.  J. Biol. Chem. (2014) 289(45):31591-604

  • Alvaro, C.G., A.F. O’Donnell, D.C. Prosser, A.A. Augustine, A. Goldman, J.L. Brodsky, M.S. Cyert, B. Wendland, and J. Thorner.  Specific a-arrestins negatively regulate Saccharomyces cerevisiae pheromone response by down-modulating the G-protein coupled receptor Ste2.  Mol. Cell. Biol. (2014) 34:2660-81.

  • Adam, C., A. Baeurle, J.L. Brodsky, P. Wipf, D. Schrama, J.C. Becker, and R. Houben.  The HSP70 Modulator MAL3-101 inhibits Merkel Cell Carcinoma. PLoS ONE (2014) 9: e92041

  • Zacchi, L.F., H.C. Wu, S.L. Bell, L. Millen, A.W. Paton, J.C. Paton, P.J. Thomas, M. Zolkiewski, and J.L. Brodsky. The BiP molecular chaperone plays multiple roles during the biogenesis of TorsinA, a AAA+ ATPase associated with the neurological disease Primary Torsion Dystonia.  J. Biol. Chem. (2014) 289: 12727-12747

  • Nakatsukasa, K., T. Kamura, and J. L. Brodsky.  Recent technical developments in the study of ER-associated degradation.  Curr. Opin. Cell Biol. (2014) 29:82–91

  • Brodsky, J.L., A. Merz, and T. Serio. Organelle and Proteome Quality Control Mechanisms: How Cells are able to Keep Calm and Carry On.  Mol. Biol. Cell (2014) 25: 733-734

  • Hecht, K.A., A.F. O’Donnell, and J.L. Brodsky. The proteolytic landscape of the yeast vacuole.  Cell. Logistics (2014) 4(1):e28023

  • Kang, Y., T. Taldone, H.J. Patel, P.D. Patel, A. Rodina, A. Gozman, R. Maharaj, C.C. Clement, M.R. Patel, J.L. Brodsky, J.C. Young, G. Chiosis. Heat shock protein 70 inhibitors. 1. 2,5'-thiodipyrimidine and 5-(phenylthio)pyrimidine acrylamides as irreversible binders to an allosteric site on heat shock protein 70.  J. Med. Chem. (2014) 57:1188-1207.

  • Brodsky, J.L. The threads that tie protein-folding diseases.  Dis. Model. Mech. (2014) 7: 3-4

  • Tran, J. R. and J. L. Brodsky.  The Cdc48-Vms1 complex maintains 26S proteasome architecture.  Biochem. J. (2014) 458: 459-467

  • Kolb, A. R., P. G. Needham, C. Rothenberg, C. J. Guerriero, P. A. Welling, and J. L. Brodsky.  ESCRT regulates surface expression of the Kir2.1 potassium channel.  Mol. Biol. Cell (2014) 25: 276- 289

  • Butkinaree, C., L. Guo, B. Ramkhelawon, A. Wanschel, J.L. Brodsky, K.J. Moore, and E.A. Fisher. A Regulator of Secretory Vesicle Size, KLHL12, Facilitates the Secretion of Apolipoprotein B100 and Very Low Density Lipoproteins.  Arterioscler. Thromb. Vasc. Biol. (2014) 34: 251-254

  • Simon, N., M. L. Bochman, S. Seguin, J. L. Brodsky, W. L. Seibel, and A. Schwacha. Ciprofloxacin is an inhibitor of the MCM2-7 replicative helicase.  Biosci. Rep. (2013) 33(5), art:e00072

  • Brodsky, J.L.  Just a trim, please: Refining ER degradation through deubiquitination.  Cell (2013) 154: 479-481

  • Guerriero, C. J., K. F. Weiberth, and J. L. Brodsky.  Hsp70 Targets a Cytoplasmic Quality Control Substrate to the San1p Ubiquitin Ligase.  J. Biol. Chem. (2013) 288: 18506-18520

  • Buck, T. M., L. Plavchak, A. Roy, B. F. Donnelly, O. B. Kashlan, T. R. Kleyman, A. R. Subramanya, J. L. Brodsky. The Lhs1/GRP170 chaperones facilitate the endoplasmic reticulum associated degradation of the epithelial sodium channel.  J. Biol. Chem. (2013) 288: 18366-18380

  • Neal, M.D., J. Hongpeng, B. Eyer, M. Good, C. J. Guerriero, C. P. Sodhi, A. Afrazi, T. Prindle Jr, C. Ma, M. Branca, J. Ozoleck, T. R. Billiar, J. L. Brodsky, P. Wipf, and D. J. Hackam.  Discovery and Validation of a New Class of Small Molecule TLR4 Inhibitors that Attenuate Endotoxemia and Necrotizing Enterocolitis.  PLoS ONE (2013) 8 (6): e65779

  • Hecht, K.A., V. Wytiaz, T. Ast, M. Schuldiner, and J.L. Brodsky.  Characterization of an M28 family member residing in the yeast vacuole.  FEMS Yeast Res. (2013) 13: 471-484

  • Kilpatrick K, Novoa JA, Hancock T, Guerriero CJ, Wipf P, Brodsky JL, Segatori L. Chemical induction of Hsp70 reduces α-synuclein aggregation in neuroglioma cells. ACS Chem Biol. (2013) 8: 1460-1468

  • Needham PG, Brodsky JL. How early studies on secreted and membrane protein quality control gave rise to the ER associated degradation (ERAD) pathway: The early history of ERAD. Biochim Biophys Acta. (2013) 1883: 2447-2457

  • Nakatsukasa K, Brodsky JL, Kamura T. A stalled retrotranslocation complex reveals physical linkage between substrate recognition and proteasomal degradation during ER associated degradation. Mol Biol Cell. (2013) 24: 1765-1775

  • Andreo U, Guo L, Chirieac DV, Tuyama AC, Montenont E, Brodsky JL, Fisher EA. Insulin-Stimulated Degradation of Apolipoprotein B100: Roles of Class II Phosphatidylinositol-3-Kinase and Autophagy. PLoS One. 2013;8(3):e57590.

  • Abisambra J, Jinwal UK, Miyata Y, Rogers J, Blair L, Li X, Seguin SP, Wang L, Jin Y, Bacon J, Brady S, Cockman M, Guidi C, Zhang J, Koren J, Young ZT, Atkins CA, Zhang B, Lawson LY, Weeber EJ, Brodsky JL, Gestwicki JE, Dickey CA. Allosteric Heat Shock Protein 70 Inhibitors Rapidly Rescue Synaptic Plasticity Deficits by Reducing Aberrant Tau. Biol Psychiatry. (2013) 74: 367-374

  • Donnelly BF, Needham PG, Snyder AC, Roy A, Khadem S, Brodsky JL, Subramanya AR. Hsp70 and Hsp90 Multichaperone Complexes Sequentially Regulate Thiazide-Sensitive Cotransporter ER-Associated Degradation and Biogenesis. J Biol Chem. (2013) 288: 13124-13135

  • Miyata Y, Li X, Lee HF, Jinwal UK, Srinivasan SR, Seguin SP, Young ZT, Brodsky JL, Dickey CA, Sun D, Gestwicki JE. Synthesis and Initial Evaluation of YM-08, a Blood-Brain Barrier Permeable Derivative of the Heat Shock Protein 70 (Hsp70) Inhibitor MKT-077, Which Reduces Tau Levels. ACS Chem Neurosci. (2013) 4: 930-939

  • Brodsky, J.L. Cleaning Up: ER-Associated Degradation to the Rescue. Cell (2012) 151: 1163-1167

  • Gelling, C. L., I. W. Dawes, D. H. Perlmutter, E. A. Fisher, and J. L. Brodsky.  The endosomal protein sorting receptor sortilin has a role in trafficking alpha-1 antitrypsin.  Genetics (2012) 192: 889-903

  • Fisher, E. A. and J. L. Brodsky.  The unfolded protein response: A multifaceted regulator of lipid and lipoprotein metabolism.  Cell Metab. (2012) 16: 407-408

  • Seguin, S. P., A. W. Ireland, T. Gupta, Y. Miyata, P. Wipf, J. M. Pipas, J. E. Gestwicki, and J. L. Brodsky.  A screen for modulators of large T antigen’s ATPase activity uncovers novel inhibitors of simian virus 40 and BK virus replication. Antiviral Res. (2012) 96: 70-81

  • Tran, J., and J. L. Brodsky.  Assays to measure ER associated degradation (ERAD) in yeast, in “Ubiquitin family modifiers and the proteasome: Reviews and protocols” (Dohmen, J., and Scheffner, M., Eds.).  Methods Mol. Biol. (2012) 832: 505-518

  • Hutt, D, M., D. M. Roth, M. Chalfant, R. T. Youker, J. Matteson, J. L. Brodsky, and W. E. Balch.  (2012) FKBP8 peptidyl-prolyl isomerase activity manages a late stage of CFTR folding and stability.  J. Biol. Chem. (2012) 287: 21914-21925

  • Arora, R., M. Shuda, A. Gustafierro, H. Feng, T. Toptan, Y. Tolstov, D. Normolle, L. L. Vollmer, A. Vogt, A. Dömling, J. L. Brodsky, Y. Chang, and P. S. Moore.  (2012) Viral knockdown reveals surviving as a therapeutic target for Merkel cell carcinoma.  Science Transl. Med. (2012) 4(133):133ra56

  • Grubb, S., L. Guo, E. A. Fisher, and J. L. Brodsky. (2012) Protein Disulfide Isomerases Contribute Differentially to the Endoplasmic Reticulum – Associated Degradation of Apolipoprotein B and Other Substrates.  Mol. Biol. Cell (2012) 23(4):520-32

  • Guerriero, C., and J. L. Brodsky. (2012) On the precipice: the delicate balance between secreted protein folding and endoplasmic reticulum-associated degradation (ERAD) in human physiology. Physiol. Rev. (2012) 92: 537-576

  • Needham, P. G., K. Mikoluk, P. Dhakarwal, S. Khadem, A. C. Snyder, A. R. Subramanya, and J. L. Brodsky. (2011)  The Thiazide-Sensitive NaCl Cotransporter is Targeted for Chaperone-Dependent ER-Associated Degradation. J. Biol. Chem286:43611-21

  • Daghestani, H.N., G. Zhu, P.A. Johnston, S. Shinde, J.L. Brodsky, and B.W. Day.  (2011)  Characterization of inhibitors of glucocorticoid receptor nuclear translocation: A model of cytoplasmic dynein-mediated cargo transport. Assay Drug Devel. Technol10:46-60

  • Braunstein, M. J., S. S. Scott, C. M. Scott, S. Behrman, P. Walter, P. Wipf, J. D. Coplan, W. J. Chirico, D. Joseph, J. L. Brodsky, and O. Batuman.  (2011)  Cytotoxic effects of the Hsp70 molecular chaperone inhibitor MAL3-101 on multiple myeloma.  J. Oncol. 2011:232037

  • Seguin, S. P., C. W. Evans, M. Nebane, S. McKellip, S. Ananthan, N. A. Tower, M. Sosa, L. Rasmussen, E. L. White, B. E. Maki, D. S. Matharu, J. E. Golden, J. Aubé, J. L. Brodsky, and J. W. Noah. (2011)  High-throughput Screening Identifies A Bisphenol Inhibitor of SV40 Large T-Antigen ATPase Activity. J. Biomol. Screen17:194-203

  • Falcone, D., M. P. Henderson, H. Nieuwland, C. M. Coughlan, J. L. Brodsky, and D. W. Andrews.  (2011)  Stability and function of the Sec61 translocation complex depends on the Sss1p tail-anchor sequence.  Biochem. J. 436: 291-303

  • Brodsky, J. L., and W. R. Skach. (2011) Protein folding and quality control in the endoplasmic reticulum: recent lessons from yeast and mammalian systems.  Curr. Opin. Cell Biol. 23: 464-475

  • Huryn, D.M., J.L. Brodsky, K.M. Brummond, P. G. Chambers, B. Eyre, A.W. Ireland, M. Kawasumi, M.G. LaPorte, K. Lloyd, B. Manteau, P. Nghiem, B. Quade, S.P. Seguin, and P. Wipf.  (2011)  Chemical Methodology as a Source of Small Molecule Checkpoint Inhibitors and Hsp70 Modulators.  Proc. Natl. Acad. Sci. USA 108: 6757-6762

  • Bell, S. L., A. Chiang, and J. L. Brodsky. (2011). A malarial Hsp70 rescues chaperone-dependent activities in ssa1 mutant yeast. PLoS One 6: e20047

  • Kolb, A. R., T. M. Buck, and J. L. Brodsky (2011) Saccharomyces cerevisiae as a model system for kidney disease: What can yeast tell us about renal function? Am. J. Physiol. 301: F1-F11

  • Yi, S.X., J.B. Benoit, M.A. Elnitsky, N. Kaufmann, J.L. Brodsky, M.L. Zeidel, D.L. Denlinger, and J.r Lee RE (2011) Function and immuno-localization of aquaporins in the Antarctic midge Belgica antarctica. J Insect Physiol 57: 1096-1105

  • Tran, J.R., L.R. Tomsic, and J.L. Brodsky (2011) A Cdc48p-associated factor modulates endoplasmic reticulum-associated degradation, cell stress, and ubiquitinated protein homeostasis. J Biol Chem 286:5744-5755

  • Botha, M., A.N. Chiang, P.G. Needham, L.L. Stephens, H.C. Hoppe, S. Kulzer, J.M. Przyborski, K. Lingelbach, P. Wipf, J.L. Brodsky, A. Shonhai, and G.L. Blatch (2011) Plasmodium falciparum encodes a single cytosolic type I Hsp40 that functionally interacts with Hsp70 and is upregulated by heat shock. Cell Stress Chaperones 16: 389-401

  • Ahn, J.H., W. Luo, J. Kim, A. Rodina, C.C. Clement, J. Aguirre, W. Sun, Y. Kang, R. Maharaj, K. Moulick, D. Zatorska, M. Kokoszka, J.L. Brodsky, and G. Chiosis (2011) Design of a Flexible Cell-Based Assay for the Evaluation of Heat Shock Protein 70 Expression Modulators. Assay Drug Dev Technol 9: 236-246

  • Gelling, C.L., and J.L. Brodsky (2010) Mechanisms underlying the cellular clearance of antitrypsin Z: lessons from yeast expression systems. Proc Am Thorac Soc 7:363-367

  • Heo, J.M., N. Livnat-Levanon, E.B. Taylor, K.T. Jones, N. Dephoure, J. Ring, J. Xie, J.L. Brodsky, F. Madeo, S.P. Gygi, K. Ashrafi, M.H. Glickman, and J. Rutter (2010) A stress-responsive system for mitochondrial protein degradation. Mol. Cell 40:465-480

  • Brodsky, J.L. (2010) The use of in vitro assays to measure endoplasmic reticulum-associated degradation. Methods Enzymol. 470:661-679

  • Brodsky, J.L. (2010) The special delivery of a tail-anchored protein: why it pays to use a dedicated courier. Mol. Cell 40:5-7

  • Goeckeler, J.L., and J.L. Brodsky (2010) Molecular chaperones and substrate ubiquitination control the efficiency of endoplasmic reticulum-associated degradation. Diabetes Obes. Metab. 12 Sup:32-38

  • Wisen, S., E.B. Bertelsen, A.D. Thompson, S. Patury, P. Ung, L. Chang, C.G. Evans, G.M. Walter, P. Wipf, H.A. Carlson, J.L. Brodsky, E.R. Zuiderweg, and J.E. Gestwicki (2010) Binding of a small molecule at a protein-protein interface regulates the chaperone activity of Hsp70-Hsp40. ACS Chem. Biol. 5:611-622

  • Vembar, S.S., M.C. Jonikas, L.M. Hendershot, J.S. Weissman, and J.L. Brodsky (2010) J domain co-chaperone specificity defines the role of BiP during protein translocation. J. Biol. Chem. 285:22484-22494

  • Nakatsukasa, K., and J.L. Brodsky (2010) In vitro reconstitution of the selection, ubiquitination, and membrane extraction of a polytopic ERAD substrate. Methods Mol. Biol. 619:365-376

  • Buck, T.M., A.R. Kolb, C.R. Boyd, T.R. Kleyman, and J.L. Brodsky (2010) The endoplasmic reticulum-associated degradation of the epithelial sodium channel requires a unique complement of molecular chaperones. Mol Biol Cell 21:1047-1058

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