The discovery that endogenously produced H2S is a signaling molecule that mediates varied physiological effects in mammals ranging from neuromodulatory to cardioprotective, has spurred enormous recent interest in understanding its biology and in exploiting its pharmacological potential. Enzymes involved in sulfur metabolism catalyze the biogenesis of H2S and include: cystathionine -synthase (CBS), - cystathionase (CSE) and mercaptopyruvate sulfurtransferase (MST). However, despite the rising interest in H2S biochemistry, fundamental questions remain regarding regulation of its production, its mechanism of action and its destruction. In this study, we propose to address significant gaps in our understanding of the reaction mechanisms and regulation of H2S production by addressing the following specific aims. (i) Structure- function studies on H2S biogenesis will focus on elucidating the reaction mechanisms of CBS, CSE and MST and conducting a high throughput screen using a small molecule library as a platform for in vitro and ex vivo screening of activators and/or inhibitors of CBS and CSE. (ii) Regulation of CBS, CSE and MST by redox, androgens and sumoylation will be investigated to determine how these parameters modulate H2S production, affect transsulfuration flux and localization of glutathione pools during the cell cycle. The impact of the proposed studies is both fundamental (i.e., gaining insights into the operation and regulation of H2S-generating enzymes) and translational (i.e., informing therapeutic options for circumventing metabolic blockades, screening for activators/inhibitors of H2S production and characterizing testosterone regulation of the transsulfuration pathway in androgen-responsive prostate cancer).

Public Health Relevance

Homocystinuria, cystathionuria and mercaptolactate-cysteine disulfiduria are rare/orphan diseases that result from defects in three enzymes in sulfur metabolism that also contribute to biogenesis of the signaling molecule, H2S, which mediates an array of physiological effects. We will elucidate the reaction mechanisms and regulation of these enzymes, which is essential for understanding how the process is corrupted in patients, and, for informing treatment options.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL058984-15
Application #
8664907
Study Section
Macromolecular Structure and Function E Study Section (MSFE)
Program Officer
Srinivas, Pothur R
Project Start
1997-08-01
Project End
2017-05-31
Budget Start
2014-06-01
Budget End
2015-05-31
Support Year
15
Fiscal Year
2014
Total Cost
Indirect Cost
Name
University of Michigan Ann Arbor
Department
Biochemistry
Type
Schools of Medicine
DUNS #
City
Ann Arbor
State
MI
Country
United States
Zip Code
48109
Ghosh, Amiya Kumar; Mau, Theresa; O'Brien, Martin et al. (2016) Impaired autophagy activity is linked to elevated ER-stress and inflammation in aging adipose tissue. Aging (Albany NY) 8:2525-2537
Wedmann, Rudolf; Onderka, Constantin; Wei, Shengwei et al. (2016) Improved tag-switch method reveals that thioredoxin acts as depersulfidase and controls the intracellular levels of protein persulfidation. Chem Sci 7:3414-3426
Carballal, Sebastián; Cuevasanta, Ernesto; Yadav, Pramod K et al. (2016) Kinetics of Nitrite Reduction and Peroxynitrite Formation by Ferrous Heme in Human Cystathionine β-Synthase. J Biol Chem 291:8004-13
Kabil, Omer; Yadav, Vinita; Banerjee, Ruma (2016) Heme-dependent Metabolite Switching Regulates H2S Synthesis in Response to Endoplasmic Reticulum (ER) Stress. J Biol Chem 291:16418-23
Yadav, Pramod K; Martinov, Michael; Vitvitsky, Victor et al. (2016) Biosynthesis and Reactivity of Cysteine Persulfides in Signaling. J Am Chem Soc 138:289-99
Banerjee, Ruma (2016) Introduction to the Thematic Minireview Series on Intrinsically Disordered Proteins. J Biol Chem 291:6679-80
Niu, Wei-Ning; Yadav, Pramod Kumar; Adamec, Jiri et al. (2015) S-glutathionylation enhances human cystathionine β-synthase activity under oxidative stress conditions. Antioxid Redox Signal 22:350-61
Vitvitsky, Victor; Banerjee, Ruma (2015) H2S analysis in biological samples using gas chromatography with sulfur chemiluminescence detection. Methods Enzymol 554:111-23
Mishanina, Tatiana V; Libiad, Marouane; Banerjee, Ruma (2015) Biogenesis of reactive sulfur species for signaling by hydrogen sulfide oxidation pathways. Nat Chem Biol 11:457-64
Gao, Xing-Huang; Krokowski, Dawid; Guan, Bo-Jhih et al. (2015) Quantitative H2S-mediated protein sulfhydration reveals metabolic reprogramming during the integrated stress response. Elife 4:e10067

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