High Constitutive Activity Is an Intrinsic Feature of Ghrelin Receptor Protein

Damian, Marjorie; Marie, Jacky; Leyris, Jean-Philippe; Fehrentz, Jean‐Alain; Verdié, Pascal; Martinez, Jean; Banères, Jean-Louis; Mary, Sophie

doi:10.1074/jbc.m111.288324

Cited by 134 publications

(71 citation statements)

References 69 publications

(101 reference statements)

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“…As the GHS receptor has been reported to display a ~50% constitutive activity3546, studying the molecular dynamics of the receptor in the presence of either agonist or inverse agonist represents an interesting topic8. Figure 5 shows the order parameters of the GHS receptor in the absence as well as in the presence of either the agonist ghrelin or the inverse agonist KbFwLK(Pam)-NH 2 recorded in DipShift experiments that were excited by CP at a contact time of 700 μs (A, B) or by direct 13 C excitation (C, D) both in POPC (A, C) and DMPC membranes (B, D).…”

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Expression, Functional Characterization, and Solid-State NMR Investigation of the G Protein-Coupled GHS Receptor in Bilayer Membranes

Schrottke

Kaiser

Vortmeier

et al. 2017

Sci Rep

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The expression, functional reconstitution and first NMR characterization of the human growth hormone secretagogue (GHS) receptor reconstituted into either DMPC or POPC membranes is described. The receptor was expressed in E. coli. refolded, and reconstituted into bilayer membranes. The molecule was characterized by 15N and 13C solid-state NMR spectroscopy in the absence and in the presence of its natural agonist ghrelin or an inverse agonist. Static 15N NMR spectra of the uniformly labeled receptor are indicative of axially symmetric rotational diffusion of the G protein-coupled receptor in the membrane. In addition, about 25% of the 15N sites undergo large amplitude motions giving rise to very narrow spectral components. For an initial quantitative assessment of the receptor mobility, 1H-13C dipolar coupling values, which are scaled by molecular motions, were determined quantitatively. From these values, average order parameters, reporting the motional amplitudes of the individual receptor segments can be derived. Average backbone order parameters were determined with values between 0.56 and 0.69, corresponding to average motional amplitudes of 40–50° of these segments. Differences between the receptor dynamics in DMPC or POPC membranes were within experimental error. Furthermore, agonist or inverse agonist binding only insignificantly influenced the average molecular dynamics of the receptor.

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Section: Resultsmentioning

confidence: 99%

“…Another obvious difference between individual members of the GPCR family is the constitutive activity of the GHS receptor, which is on the order of 50%3546. As well investigated for the β 2 adrenergic receptor, ligand binding alters the shape of the energy landscape and the receptor populates different conformations820.…”

Section: Discussionmentioning

confidence: 99%

Expression, Functional Characterization, and Solid-State NMR Investigation of the G Protein-Coupled GHS Receptor in Bilayer Membranes

Schrottke

Kaiser

Vortmeier

et al. 2017

Sci Rep

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“…They can be, for example, extensively diluted with APol-free buffer (Tribet et al 2009; Zoonens et al 2007), washed with surfactant-free buffer after adsorption onto solid supports (Charvolin et al 2009; Della Pia et al 2014a, b; Giusti et al 2014a; Le Bon et al 2014a), or injected on a SEC column pre-equilibrated with surfactant-free buffer (Charvolin et al 2014; Gohon et al 2008; Zoonens et al 2007). Nevertheless, the APol layer can be easily displaced and exchanged upon exposure to an excess of competing surfactants, be they detergents (Damian et al 2012; Tribet et al 1997, 2009; Zoonens et al 2007), lipid vesicles (Nagy et al 2001), black lipid membranes (BLM) (Pocanschi et al 2006), lipidic three-dimensional (3D) phases (Polovinkin et al 2014b), cell plasma membranes (Popot et al 2011), or free APols (Tribet et al 1997; Zoonens et al 2007) (Fig. 3b).…”

Section: Basic Properties Of Amphipols and Membrane Protein/amphipolmentioning

confidence: 99%

“…The latter route has been used to transfer to NDs a GPCR that had been folded in A8-35 (Damian et al 2012). …”

Section: Applicationsmentioning

confidence: 99%

Amphipols for Each Season

2014

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Amphipols (APols) are short amphipathic polymers that can substitute for detergents at the transmembrane surface of membrane proteins (MPs) and, thereby, keep them soluble in detergent-free aqueous solutions. APol-trapped MPs are, as a rule, more stable biochemically than their detergent-solubilized counterparts. APols have proven useful to produce MPs, most noticeably by assisting their folding from the denatured state obtained after solubilizing MP inclusion bodies in either SDS or urea. They facilitate the handling in aqueous solution of fragile MPs for the purpose of proteomics, structural and functional studies, and therapeutics. Because APols can be chemically labeled or functionalized, and they form very stable complexes with MPs, they can also be used to functionalize those indirectly, which opens onto many novel applications. Following a brief recall of the properties of APols and MP/APol complexes, an update is provided of recent progress in these various fields.

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“…In this regard, an interesting feature of GHSR-1a is its high constitutive activity that makes it able to signal in a ghrelin-independent manner [41,42]. Such constitutive activity of GHSR-1a has been not only shown to strongly impact on the neuronal activity but also proposed to have some physiological impact [43,44,45].…”

Section: Introductionmentioning

confidence: 99%

Is Ghrelin Synthesized in the Central Nervous System?

Cabral

Soto

Epelbaum

et al. 2017

IJMS

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Abstract:Ghrelin is an octanoylated peptide that acts via its specific receptor, the growth hormone secretagogue receptor type 1a (GHSR-1a), and regulates a vast variety of physiological functions. It is well established that ghrelin is predominantly synthesized by a distinct population of endocrine cells located within the gastric oxyntic mucosa. In addition, some studies have reported that ghrelin could also be synthesized in some brain regions, such as the hypothalamus. However, evidences of neuronal production of ghrelin have been inconsistent and, as a consequence, it is still as a matter of debate if ghrelin can be centrally produced. Here, we provide a comprehensive review and discussion of the data supporting, or not, the notion that the mammalian central nervous system can synthetize ghrelin. We conclude that no irrefutable and reproducible evidence exists supporting the notion that ghrelin is synthetized, at physiologically relevant levels, in the central nervous system of adult mammals.

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High Constitutive Activity Is an Intrinsic Feature of Ghrelin Receptor Protein

Cited by 134 publications

References 69 publications

Expression, Functional Characterization, and Solid-State NMR Investigation of the G Protein-Coupled GHS Receptor in Bilayer Membranes

Expression, Functional Characterization, and Solid-State NMR Investigation of the G Protein-Coupled GHS Receptor in Bilayer Membranes

Amphipols for Each Season

Is Ghrelin Synthesized in the Central Nervous System?

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