Patrick Thibodeau Ph.D.

Assistant Professor

Training: University of Texas at Austin, B,A.: Zoology, 1998 University of Texas Southwestern Medical Center at Dallas, Ph.D.: Molecular Biophysics, 2006 Philip Thomas Laboratory, CFTR Structure and Folding University of Texas Southwestern Medical Center at Dallas, Postdoctoral Associate, 2006-2007 Philip Thomas Laboratory, CFTR Structure and Folding Personal Statement I am particularly interested in understanding how protein folding and dynamics influence physiological functions, impacting human health and disease. The regulation of protein folding and dynamics is fundamental for all proteins and is incompletely understood. This work has focused on protein folding events that regulate epithelial physiology, either from within the epithelial (host) cell or from pathogens that act to alter epithelial function. My training is in protein biophysics and structure, with an emphasis on protein folding and dynamics. We seek to apply biophysical and structural approaches to inform our understanding of pathophysiology and disease. I have been fortunate to train in exceptional environments and with excellent biophysicists and utilize this training to support the training of students and post-doctoral associates on our campus. My laboratory in Pittsburgh has focused on furthering these studies on membrane proteins, with en emphasis on cystic fibrosis related disease processes. This work has extended studies on human ABC transporters and ion channels and developed to explore how the same fundamental protein dynamics questions regulate host-pathogen interactions. At the heart of these studies, we are interested in understanding the basic structural and functional defects associated with human disease processes. These studies focus on developing our understanding of the earliest steps in which mutations impact proteins – during early steps in biogenesis and folding – and the consequences of altered protein structure on function and physiology. Positions, Professional Experience and Honors 2014- Assistant Professor, Dept. of Microbiology and Molecular Genetics, University of Pittsburgh School of Medicine 2007-14 Assistant Professor, Dept. of Cell Biology and Molecular Physiology, University of Pittsburgh School of Medicine 2006-7 University of Texas Southwestern Medical Center at Dallas, Post-doctoral Fellow 2000-6 NIH Predoctoral Training Grant Recipient 1996-7 University of Texas at Austin Dean's Honor List Contributions to Science 1. Folding and structure of CFTR and human ABC transporters My doctoral and post-doctoral training began with a focus on human ABC transporters with a goal of understanding how disease-causing mutations impacted protein biosynthesis, folding and trafficking. These studies have continued in my laboratory at the University of Pittsburgh with the purpose of developing more complete models of ABC transporter folding pathways, identifying critical mechanisms of biosynthetic regulation, and elucidating mechanisms of functional regulation. These studies have focused on CFTR, the protein associated with cystic fibrosis, and a closely related human protein, ABCC6, which is associated with pseudoxanthoma elasticum. These studies have lead to the elucidation of a multi-step biosynthetic pathway and mechanism that facilitate suppression of folding and trafficking defects associated with disease-causing mutations. Thibodeau, P.H., Brautigam, C.A., Machius, M., Thomas, P.J. (2005) Side chain and backbone contributions of Phe508 to CFTR folding. Nat Struct Mol Biol 12:10-16. Thibodeau, P.H., Richardson M, Wang W, Millen L, Watson J, Mendoza JL, Du K, Fischman S, Senderowitz H, Lukacs GL, Thomas PJ. (2010) The cystic fibrosis-causing mutation {Delta}F508 affects multiple steps in CFTR biogenesis. JBC. 285: 35825-35835. Xue P, Crum CM, and Thibodeau PH. (2014) Regulation of ABCC6 biosynthesis and trafficking by a conserved C-terminal PDZ-like sequence. PLoSOne. 9(5):e97360. Ran, Y and Thibodeau PH. (2017) Stabilization of Nucleotide Binding Domain Dimers Rescues ABCC6 Mutants Associated with Pseudoxanthoma Elasticum. JBC. 292(5): 1559-1572. 2. Novel modes of serralysin virulence and cytotoxicity in epithelial cells Our initial studies on the serralysin protease semerged from suggestions that extracellular proteases may cleave the epithelial sodium channel (ENaC), causing tis activation. This in turn would putatively lead to the absorption of sodium and dehydration of airway surface liquid in the lung. As result, this proteolytic activation would decrease mucocilliary clearance rates and facilitate bacterial adhesion and colonization. Our studies began with understanding the funcational regulation of the alkaline protease (ArpA) from Pseudomonas aeruginosa and its role ability to activate ENaC. These studies showed that the protease was active and capable of cleaving ENaC in airway cells. These studies showed, for the first time, that a metalloprotease was capable of activating ENaC and that this mode of activation may be utilized by pathogens to modulate host defenses. We have extended these studies with the shanks laboratory to explore serralysin proteases from Serratia and their role in modulating virulence in airway and ocular cells. Zhang L, and Thibodeau PH. (2012) Calcium induced folding and stabilization of the Pseudomonas aeruginosa alkaline protease. JBC. 287:4611-4322. Butterworth MB, Zhang L, Heidrich H, Myerburg MM, and Thibodeau PH, (2012) Activation of the epithelial sodium channel (ENaC) by the alkaline protease from Pseudomonas aeruginosa. J. Biol. Chem. 287(39):32556-65. Butterworth MB, Zhang L, Liu X, Shanks RMQ, Thibodeau PH. (2014) Regulation of ENaC activity by bacterial metalloproteases and inhibitors. PLoSOne. 9(6):e100313. Shanks RMQ, Stella NA, Hunt KM, Brothers KM, Zhang L, and Thibodeau PH. (2015) Identification of SlpB, a cytotoxic protease from Serratia marcescens. Infection and Immunity. Jul; 82(7): 2907-16. 3. Folding and dynamics of serralysin proteases Studies of the serralysin proteases have been developed to understand their conformational plasticity, structural stability and functional regulation. these protease are important virulence factors in multiple disease states and have unique structural properties that contribute to their virulence. We initially characterized the folding pathway of the AprA protease from Pseudomonas, showing that Ca2+ regulated the folding and activation of the protease. These studies provided key insights into RTX protein folding and the dynamics associated with Ca2+ binding. In addition, biophysical and functional studies have elucidated the interactions that regulate their stability and non- native conformations. These studies inform our understanding of RTX protein folding and have direct implications for the structural and secretions studies proposed in our application. Zhang L, and Thibodeau PH. (2012) Calcium induced folding and stabilization of the Pseudomonas aeruginosa alkaline protease. JBC. 287:4611-4322. Zhang L, Conway J and Thibodeau PH. (2014) Inducible polymerization and two-dimensional assembly of the Repeats-in-Toxin (RTX) domain from the Pseudomonas aeruginosa alkaline protease. Biochemistry. 53(41):6452-62. Shanks RMQ, Stella NA, Hunt KM, Brothers KM, Zhang L, and Thibodeau PH. (2015) Identification of SlpB, a cytotoxic protease from Serratia marcescens. Infection and Immunity. Jul; 82(7): 2907-16. Zhang L, Morrison AJ, Thibodeau PH. (2015) Interdomain Contacts and the Stability of Serralysin Protease from Serratia marcescens. PLoS One. Sep 17; 10(9):e0138419 Complete List of Published Work in PubMed: http://www.ncbi.nlm.nih.gov/pu... D. Research Support Ongoing Research Support R01GM12291 (P.i.: Thibodeau P.H.) 09/17-08/21 NIH/NIGMS Altered biosynthesis and function of ABCC6 in systemic mineralization disorders THIBOD09160X (P.I.: Thibodeau P.H.) 06/09-09/18 Cystic Fibrosis Foundation (CFF/CFFT) STRUCTURAL INTERACTIONS REGULATING CFTR FUNCTION R01EY027331 (P.I. Shanks, R.M.Q.) 04/14-03/21 NIH/NEI BACTERIAL FACTORS THAT REGULATE OCULAR HOST-PATHOGEN INTERACTIONS AND MICROBIAL KERATITIS - Consultant R01DK068196 (P.I.: Frizzell, R.A.) 05/14-04/18 NIH/NIDDK CHAPERONE INTERACTIONS IN CFTR BIOSYNTHESIS - Consultant Completed Research Support R56DK083284 (P.I.: Thibodeau, P.H.) 5/1/10-4/30/11 NIH/NIDDK REGULATED BIOSYNTHESIS AND FUNCTION OF ABC-TRANSPORT SYSTEMS R01DK083284 (P.I.: Thibodeau, P.H.) NIH/NIDDK REGULATED BIOSYNTHESIS AND FUNCTION OF ABC-TRANSPORT SYSTEMS


Appearances