Endothelin-converting enzyme 1 degrades neuropeptides in endosomes to control receptor recycling

Roosterman, Dirk; Cottrell, Graeme S.; Padilla, Benjamin E.; Muller, Laurent; Eckman, Christopher B.; Bunnett, Nigel W.; Steinhoff, Martin

doi:10.1073/pnas.0701910104

Cited by 69 publications

(165 citation statements)

References 35 publications

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“…PD-069185 (19) was a gift (Pfizer). Other reagents were from Sigma or have been described (12,13,20).…”

Section: Methodsmentioning

confidence: 99%

“…Transfected Cells and Cell Lines-Generation and maintenance of human embryonic kidney 293 (HEK) FLP cells (Invitrogen) and HEKFLP stably expressing rat NK 1 R have been described (12,13,20). In some experiments cells were transiently transfected with rat NK 1 R, rat FLAG-NK 1 R⌬325-407, rat ␤-arrestin1 V53D, rat ␤-arrestin1 319 -418 , human ␤-arrestin2-GFP, human ECE-1c-GFP, or empty vector (control) using Lipofectamine2000 (Invitrogen).…”

Section: Methodsmentioning

confidence: 99%

“…Membranes were incubated with antibodies to pERK2 (E-4; 1:1,000), ERK2 (C-14; 1:5,000), c-Fos (9F6, 1:2,000), ECE-1 (1:1,000), EEA1 (1:10,000), Rab5a (1:2,000), LAMP1 (1:2,000), Na ϩ -K ϩ -ATPase (1:5,000), histone H3 (1:20,000), NK 1 R (1:10,000), ␤-arrestin2 (1:5,000), Src (1:1,000), Nur77 (1:200), phosphothreonine (1:1,000), or ␤-actin (A-5441, 1:10,000) overnight at 4°C. Membranes were treated with secondary or tertiary antibodies and analyzed with an Odyssey Infrared Imaging System (Li-COR Biosciences) (12,13,20).…”

Section: Methodsmentioning

confidence: 99%

“…We hypothesized that mechanisms that regulate GPCRs at the plasma membrane also control receptor signaling and trafficking at the endosomal membrane. We recently reported that the metalloendopeptidase endothelin-converting enzyme-1 (ECE-1) is present at the endosomal membrane and that ECE-1 degrades internalized neuropeptides within acidified early endosomes (12)(13)(14). This degradation disrupts the peptide-GPCR-␤-arrestin complex to promote ␤-arrestin translocation back to the cytosol and receptor recycling to the cell-surface, which mediates resensitization.…”

mentioning

confidence: 99%

“…SP is expressed in the nervous and immune systems and interacts with the neurokinin 1 receptor (NK 1 R) to control neurogenic inflammation, pain, neurodegeneration, smooth muscle contraction, and exocrine secretions (15). SP induces NK 1 R interaction with ␤-arrestins at the plasma membrane, which mediates desensitization and causes ␤-arrestin-dependent endocytosis of the SP-NK 1 R-␤-arrestin complex to early endosomes containing ECE-1 (13,16). ␤-Arrestins recruit Src, MEK, and ERKs to the NK 1 R at the endosomal membrane, which mediates the effects of SP on gene expression and cell survival (6).…”

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confidence: 99%

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Endosomal Endothelin-converting Enzyme-1

Cottrell

Padilla

Amadesi

et al. 2009

Journal of Biological Chemistry

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Neuropeptide signaling at the cell surface is regulated by metalloendopeptidases, which degrade peptides in the extracellular fluid, and ␤-arrestins, which interact with G protein-coupled receptors (GPCRs) to mediate desensitization. ␤-Arrestins also recruit GPCRs and mitogen-activated protein kinases to endosomes to allow internalized receptors to continue signaling, but the mechanisms regulating endosomal signaling are unknown. We report that endothelin-converting enzyme-1 (ECE-1) degrades substance P (SP) in early endosomes of epithelial cells and neurons to destabilize the endosomal mitogen-activated protein kinase signalosome and terminate signaling. ECE-1 inhibition caused endosomal retention of the SP neurokinin 1 receptor, ␤-arrestins, and Src, resulting in markedly sustained ERK2 activation in the cytosol and nucleus, whereas ECE-1 overexpression attenuated ERK2 activation. ECE-1 inhibition also enhanced SP-induced expression and phosphorylation of the nuclear death receptor Nur77, resulting in cell death. Thus, endosomal ECE-1 attenuates ERK2-mediated SP signaling in the nucleus to prevent cell death. We propose that agonist availability in endosomes, here regulated by ECE-1, controls ␤-arrestin-dependent signaling of endocytosed GPCRs.Signal transduction at the plasma membrane is tightly regulated by well characterized mechanisms. For the large family of G protein-coupled receptors (GPCRs), 2 regulation of extracellular agonist concentration and receptor coupling to heterotrimeric G proteins control signal transduction. Cell-surface metalloendopeptidases, exemplified by neprilysin, degrade neuropeptides in the extracellular fluid to regulate receptor activation (1, 2). G protein receptor kinases phosphorylate activated GPCRs to promote their interaction with ␤-arrestins, which uncouple receptors from G proteins and mediate desensitization (3). However, activated GPCRs internalize, and endocytosed receptors continue to signal by G protein-independent mechanisms. ␤-Arrestins mediate endocytosis and intracellular signaling of GPCRs. ␤-Arrestins couple GPCRs to clathrin and AP2 to mediate endocytosis (4, 5) and are scaffolds that recruit Src, Raf-1, and mitogen-activated protein kinases (MAPKs) to GPCRs in endosomes forming a MAPK signalosome, which determines the location and function of activated extracellular signal-regulated kinases (ERKs) (6 -9). Compared with our understanding of receptor regulation at the plasma membrane, little is known about the mechanisms that regulate GPCR signaling and trafficking at the endosomal membrane. Specifically, it is not known whether agonist interaction with GPCRs or GPCR interaction with ␤-arrestins is necessary for receptor signaling in endosomes. Regulation of these interactions could determine the duration of ␤-arrestin-mediated ERK activation, with important functional implications (10, 11). We hypothesized that mechanisms that regulate GPCRs at the plasma membrane also control receptor signaling and trafficking at the endosomal membrane. We recently reported t...

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“…PD-069185 (19) was a gift (Pfizer). Other reagents were from Sigma or have been described (12,13,20).…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Endosomal Endothelin-converting Enzyme-1

Cottrell

Padilla

Amadesi

et al. 2009

Journal of Biological Chemistry

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Inflammation, Immunology and Allergy

Steinhoff

2016

Rook's Textbook of Dermatology, Ninth Edition

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Inflammation can be defined as a complex body defence mechanism responding to dangerous endogenous or exogenous stimuli in order to restore body homeostasis. Thus, the skin is continuously challenged to prevent inflammation, and inflammatory skin diseases are very common worldwide. Similarly, allergic reactions (types 1–4) are inflammatory processes created by the body system to respond to ‘danger signals’ of various origins. Very early in phylogenesis, inflammation is associated with the host defence against infectious, allergic or toxic agents, UV radiation, noxious heat and cold, or other injury. Innate and later adaptive immune mechanisms have been established that, uncontrolled, potentially lead to chronic disease or death, as in anaphylactic shock. The body system responds with the induction of various inflammatory pathways such as radical oxygen species, nitric oxide derivatives, complement compounds, adenosine monophosphate, antimicrobial peptides, cytokines, chemokines, prostaglandins, leukotrienes, proteases, neuropeptides and growth factors, just to name a few. This chapter systematically covers the various mechanisms that regulate inflammatory and allergic responses and refers in a translational setting to the diseases that are involved in order better to understand the clinical characteristics of a skin disease, its differential diagnoses and optimal treatment options.

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