A wide variety of proteins in neuronal, endocrine and immune tissues undergo proteolytic processing. Many of these proteins and peptides are intercellular messengers. Most neuroendocrine peptides are synthesized from precursor proteins. Post-translational processing of these precursors is a key step in the production of biologically active peptides. In the majority of cases this occurs by proteolysis of the precursors at 'classical'cleavage sites involving endopeptidases of the subtilisin family. A subset of bioactive peptides are generated by processing at 'non-classical'cleavage sites;the protease(s) responsible for these cleavages have not been well explored. Members of the metalloendoprotease family are thought to be involved in non- classical processing. Among them, endothelin converting enzyme-2 (ECE-2) is a good candidate since it exhibits functional properties that are consistent with it being a neuropeptide biosynthetic enzyme. In this grant application we propose to examine the hypothesis that """"""""ECE-2 is neuropeptide processing enzyme that generates novel bioactive peptides by non-classical processing"""""""".
The specific aims are, (i) to characterize the biochemical and enzymological properties of ECE-2, (ii) to study the behavioral phenotype of animals lacking ECE-2 including addictive behaviors, and (iii) to examine the peptide profile in mice lacking ECE-2 using differential isotopic labeling and mass spectrometry. These studies are synergistic and will expand our current knowledge of neuropeptide processing by providing information about novel neuropeptides. Furthermore, findings from these studies will form a basis for future research exploring the functions of these novel neuropeptides in vivo. Since a variety of neuropeptides are involved in normal functioning of the brain, understanding the mechanisms that regulate their endogenous levels becomes a prerequisite to the development of strategies for the treatment of a variety of pathologies including neurodegenerative disorders and drug addiction.

Public Health Relevance

This proposal describes studies on endothelin converting enzyme-2 - a peptide processing enzyme shown to be involved in a number of disorders including drug addiction and Alzheimer's disease. The findings from the proposed studies would help in the development of novel therapeutics for the treatment of these and other neuroendocrine disorders.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS026880-23
Application #
8456161
Study Section
Molecular Neuropharmacology and Signaling Study Section (MNPS)
Program Officer
Gnadt, James W
Project Start
1989-08-01
Project End
2014-04-30
Budget Start
2013-05-01
Budget End
2014-04-30
Support Year
23
Fiscal Year
2013
Total Cost
$342,124
Indirect Cost
$140,281
Name
Icahn School of Medicine at Mount Sinai
Department
Pharmacology
Type
Schools of Medicine
DUNS #
078861598
City
New York
State
NY
Country
United States
Zip Code
10029
Gupta, Achla; Gomes, Ivone; Bobeck, Erin N et al. (2016) Collybolide is a novel biased agonist of κ-opioid receptors with potent antipruritic activity. Proc Natl Acad Sci U S A 113:6041-6
Wardman, Jonathan H; Gomes, Ivone; Bobeck, Erin N et al. (2016) Identification of a small-molecule ligand that activates the neuropeptide receptor GPR171 and increases food intake. Sci Signal 9:ra55
Gomes, Ivone; Ayoub, Mohammed Akli; Fujita, Wakako et al. (2016) G Protein-Coupled Receptor Heteromers. Annu Rev Pharmacol Toxicol 56:403-25
Pena, Darlene Aparecida; Andrade, Victor Piana de; Silva, Gabriela Ávila Fernandes et al. (2016) Rational design and validation of an anti-protein kinase C active-state specific antibody based on conformational changes. Sci Rep 6:22114
Gomes, Ivone; Bobeck, Erin N; Margolis, Elyssa B et al. (2016) Identification of GPR83 as the receptor for the neuroendocrine peptide PEN. Sci Signal 9:ra43
Stockert, Jennifer A; Devi, Lakshmi A (2015) Advancements in therapeutically targeting orphan GPCRs. Front Pharmacol 6:100
Gupta, A; Fujita, W; Gomes, I et al. (2015) Endothelin-converting enzyme 2 differentially regulates opioid receptor activity. Br J Pharmacol 172:704-19
Fujita, Wakako; Gomes, Ivone; Devi, Lakshmi A (2015) Heteromers of μ-δ opioid receptors: new pharmacology and novel therapeutic possibilities. Br J Pharmacol 172:375-87
Stockton Jr, Steven D; Gomes, Ivone; Liu, Tong et al. (2015) Morphine Regulated Synaptic Networks Revealed by Integrated Proteomics and Network Analysis. Mol Cell Proteomics 14:2564-76
Yang, Lili; Rozenfeld, Raphael; Wu, Defeng et al. (2014) Cannabidiol protects liver from binge alcohol-induced steatosis by mechanisms including inhibition of oxidative stress and increase in autophagy. Free Radic Biol Med 68:260-7

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