Research Departments & Organizations
Cellular & Molecular Physiology: Rinehart Lab
Our laboratory is focused on the discovery and characterization of novel sites of phosphorylation in kinases and regulatory networks of proteins that control electrolyte homeostasis. Our long term goal is to understand and “decode” complex signaling networks in physiological systems. Our research integrates cutting edge proteomics with mainstay techniques of molecular biology and physiology, to study signaling networks and provide critical new insight into regulated ion transport. We are focusing on mechanisms of cellular signaling transduction that involve protein phosphorylation. Many of the cellular signaling mechanisms that are disrupted in disease states, and are assumed to be fundamental for normal cellular physiology, are largely uncharacterized and hold a vast amount of therapeutic potential. Understanding the human phosphoproteome is a major challenge as research continues ever farther into the post genome era. We therefore employ established quantitative proteomic techniques, and aim to develop novel methods, in an effort to decode new signaling pathways. We aim to find new roles for kinases, phosphatases, their substrates, and protein-protein interactions on a system-wide level. Collectively, we hope these efforts will identify molecular mechanisms important for both healthy and disease states in humans.
Tens of thousands of phosphorylation sites in diverse eukaryotic proteins have been identified through large scale mass spectrometry studies. For the great majority of these sites, however, the responsible kinase is unknown, and the functional role of phosphorylation is not understood. We have recently made substantial progress towards narrowing this critical gap in knowledge with a new technology that enables site-specific incorporation of phosphoserine into proteins. This technology utilizes an E. coli strain with an expanded genetic code and contains a dedicated sense codon for phosphoserine. We use this breakthrough technology to synthesize human phosphoproteins and accelerate our efforts in “decoding” the human phosphoproteome.
Specialized Terms: Physiological Systems; Protein Phosphorylation; Cell Signaling; Phosphoproteomics; Protein Engineering; Ion Transport; Synthetic Biology; Translational Research
Learn more at http://rinehart.commons.yale.edu/
Evolution of translation machinery in recoded bacteria enables multi-site incorporation of nonstandard amino acids.
Amiram M, Haimovich AD, Fan C, Wang YS, Aerni HR, Ntai I, Moonan DW, Ma NJ, Rovner AJ, Hong SH, Kelleher NL, Goodman AL, Jewett MC, Söll D, Rinehart J, Isaacs FJ. Evolution of translation machinery in recoded bacteria enables multi-site incorporation of nonstandard amino acids. Nature Biotechnology 2015, 33:1272-1279. 2015
A flexible codon in genomically recoded Escherichia coli permits programmable protein phosphorylation.
Pirman NL, Barber KW, Ma NJ, Haimovich AD, Rogulina S, Isaacs FJ, and Rinehart J. Nature Communications. 2015. 6, 8130. 2015
Robust Production of Recombinant Phosphoproteins Using Cell-Free Protein Synthesis.
Oza JP, Aerni HR, Pirman NL, Rogulina S, ter Haar CM, Isaacs FJ, Rinehart J, and Jewett MC. Nature Communications 2015. 6, 8168 2015
The PINK1-PARKIN Mitochondrial Ubiquitylation Pathway Drives a Program of OPTN/NDP52 Recruitment and TBK1 Activation to Promote Mitophagy.
Heo JM, Ordureau A, Paulo JA, Rinehart J, Harper JW. The PINK1-PARKIN Mitochondrial Ubiquitylation Pathway Drives a Program of OPTN/NDP52 Recruitment and TBK1 Activation to Promote Mitophagy. Molecular Cell 2015, 60:7-20. 2015
Genomically recoded organisms expand biological functions.
Lajoie MJ, Rovner AJ, Goodman DB, Aerni HR, Haimovich AD, Kuznetsov G, Mercer JA, Wang HH, Carr PA, Mosberg JA, Rohland N, Schultz PG, Jacobson JM, Rinehart J, Church GM, Isaacs FJ. Genomically recoded organisms expand biological functions. Science (New York, N.Y.) 2013, 342:357-60. 2013
Expanding the genetic code of Escherichia coli with phosphoserine.
Park HS, Hohn MJ, Umehara T, Guo LT, Osborne EM, Benner J, Noren CJ, Rinehart J, Söll D. Expanding the genetic code of Escherichia coli with phosphoserine. Science (New York, N.Y.) 2011, 333:1151-4. 2011
Sites of regulated phosphorylation that control K-Cl cotransporter activity.
Rinehart J, Maksimova YD, Tanis JE, Stone KL, Hodson CA, Zhang J, Risinger M, Pan W, Wu D, Colangelo CM, Forbush B, Joiner CH, Gulcicek EE, Gallagher PG, Lifton RP. Sites of regulated phosphorylation that control K-Cl cotransporter activity. Cell 2009, 138:525-36. 2009
Mineralocorticoid receptor phosphorylation regulates ligand binding and renal response to volume depletion and hyperkalemia.
Shibata S, Rinehart J, Zhang J, Moeckel G, Castañeda-Bueno M, Stiegler AL, Boggon TJ, Gamba G, Lifton RP. Mineralocorticoid receptor phosphorylation regulates ligand binding and renal response to volume depletion and hyperkalemia. Cell Metabolism 2013, 18:660-71. 2013