Regulation of ghrelin structure and membrane binding by phosphorylation

Eva Dehlin; Jianhua Liu; Samuel H. Yun; Elizabeth Fox; Sandra Snyder; Cyrille Gineste; Leslie Willingham; Mario Geysen; Bruce D. Gaylinn; Julianne J. Sando

doi:10.1016/j.peptides.2008.02.001

Regulation of ghrelin structure and membrane binding by phosphorylation

Eva Dehlin, Jianhua Liu, Samuel H. Yun, Elizabeth Fox, Sandra Snyder, Cyrille Gineste, Leslie Willingham, Mario Geysen, Bruce D. Gaylinn, Julianne J. Sando

Research output: Contribution to journal › Article › peer-review

22 Scopus citations

Abstract

The peptide hormone ghrelin requires Ser-3 acylation for receptor binding, orexigenic and anti-inflammatory effects. Functions of desacylghrelin are less well understood. In vitro kinase assays reveal that the evolutionarily conserved Ser-18 in the basic C-terminus is an excellent substrate for protein kinase C. Circular dichroism reveals that desacylghrelin is ∼12% helical in aqueous solution and ∼50% helical in trifluoroethanol. Ser-18-phosphorylation, Ser-18-Ala substitution, or Ser-3-acylation reduces the helical character in trifluoroethanol to ∼24%. Both ghrelin and desacylghrelin bind to phosphatidylcholine:phosphatidylserine sucrose-loaded vesicles in a phosphatidylserine-dependent manner. Phosphoghrelin and phosphodesacylghrelin show greatly diminished phosphatidylserine-dependent binding. These results are consistent with binding of ghrelin and desacylghrelin to acidic lipids via the basic face of an amphipathic helix with Ser-18 phosphorylation disrupting both helical character and membrane binding.

Original language	English (US)
Pages (from-to)	904-911
Number of pages	8
Journal	Peptides
Volume	29
Issue number	6
DOIs	https://doi.org/10.1016/j.peptides.2008.02.001
State	Published - Jun 2008
Externally published	Yes

Keywords

Ghrelin phosphorylation
Ghrelin structure
PKC substrate
Peptide-membrane binding

ASJC Scopus subject areas

Biochemistry
Physiology
Endocrinology
Cellular and Molecular Neuroscience

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10.1016/j.peptides.2008.02.001

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@article{e729c09fc301481b8b0558cb4f297803,

title = "Regulation of ghrelin structure and membrane binding by phosphorylation",

abstract = "The peptide hormone ghrelin requires Ser-3 acylation for receptor binding, orexigenic and anti-inflammatory effects. Functions of desacylghrelin are less well understood. In vitro kinase assays reveal that the evolutionarily conserved Ser-18 in the basic C-terminus is an excellent substrate for protein kinase C. Circular dichroism reveals that desacylghrelin is ∼12% helical in aqueous solution and ∼50% helical in trifluoroethanol. Ser-18-phosphorylation, Ser-18-Ala substitution, or Ser-3-acylation reduces the helical character in trifluoroethanol to ∼24%. Both ghrelin and desacylghrelin bind to phosphatidylcholine:phosphatidylserine sucrose-loaded vesicles in a phosphatidylserine-dependent manner. Phosphoghrelin and phosphodesacylghrelin show greatly diminished phosphatidylserine-dependent binding. These results are consistent with binding of ghrelin and desacylghrelin to acidic lipids via the basic face of an amphipathic helix with Ser-18 phosphorylation disrupting both helical character and membrane binding.",

keywords = "Ghrelin phosphorylation, Ghrelin structure, PKC substrate, Peptide-membrane binding",

author = "Eva Dehlin and Jianhua Liu and Yun, {Samuel H.} and Elizabeth Fox and Sandra Snyder and Cyrille Gineste and Leslie Willingham and Mario Geysen and Gaylinn, {Bruce D.} and Sando, {Julianne J.}",

note = "Funding Information: We thank Dr. Nick Sherman of the University of Virginia Biomolecular Research Facility for the ghrelin phosphopeptide analysis via mass spectrometry, Dr. David Cafiso for advice on deconvolution of CD spectra and Tamara Stoops and Vanessa King-Mangard for technical assistance at the early stages of this work. This work was supported by HHS grant GM31184 (to JJS.) and by a pilot feasibility grant from the University of Virginia Silvio O. Conte Digestive Disease Research Center. ",

year = "2008",

month = jun,

doi = "10.1016/j.peptides.2008.02.001",

language = "English (US)",

volume = "29",

pages = "904--911",

journal = "Peptides",

issn = "0196-9781",

publisher = "Elsevier Inc.",

number = "6",

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TY - JOUR

T1 - Regulation of ghrelin structure and membrane binding by phosphorylation

AU - Dehlin, Eva

AU - Liu, Jianhua

AU - Yun, Samuel H.

AU - Fox, Elizabeth

AU - Snyder, Sandra

AU - Gineste, Cyrille

AU - Willingham, Leslie

AU - Geysen, Mario

AU - Gaylinn, Bruce D.

AU - Sando, Julianne J.

N1 - Funding Information: We thank Dr. Nick Sherman of the University of Virginia Biomolecular Research Facility for the ghrelin phosphopeptide analysis via mass spectrometry, Dr. David Cafiso for advice on deconvolution of CD spectra and Tamara Stoops and Vanessa King-Mangard for technical assistance at the early stages of this work. This work was supported by HHS grant GM31184 (to JJS.) and by a pilot feasibility grant from the University of Virginia Silvio O. Conte Digestive Disease Research Center.

PY - 2008/6

Y1 - 2008/6

N2 - The peptide hormone ghrelin requires Ser-3 acylation for receptor binding, orexigenic and anti-inflammatory effects. Functions of desacylghrelin are less well understood. In vitro kinase assays reveal that the evolutionarily conserved Ser-18 in the basic C-terminus is an excellent substrate for protein kinase C. Circular dichroism reveals that desacylghrelin is ∼12% helical in aqueous solution and ∼50% helical in trifluoroethanol. Ser-18-phosphorylation, Ser-18-Ala substitution, or Ser-3-acylation reduces the helical character in trifluoroethanol to ∼24%. Both ghrelin and desacylghrelin bind to phosphatidylcholine:phosphatidylserine sucrose-loaded vesicles in a phosphatidylserine-dependent manner. Phosphoghrelin and phosphodesacylghrelin show greatly diminished phosphatidylserine-dependent binding. These results are consistent with binding of ghrelin and desacylghrelin to acidic lipids via the basic face of an amphipathic helix with Ser-18 phosphorylation disrupting both helical character and membrane binding.

AB - The peptide hormone ghrelin requires Ser-3 acylation for receptor binding, orexigenic and anti-inflammatory effects. Functions of desacylghrelin are less well understood. In vitro kinase assays reveal that the evolutionarily conserved Ser-18 in the basic C-terminus is an excellent substrate for protein kinase C. Circular dichroism reveals that desacylghrelin is ∼12% helical in aqueous solution and ∼50% helical in trifluoroethanol. Ser-18-phosphorylation, Ser-18-Ala substitution, or Ser-3-acylation reduces the helical character in trifluoroethanol to ∼24%. Both ghrelin and desacylghrelin bind to phosphatidylcholine:phosphatidylserine sucrose-loaded vesicles in a phosphatidylserine-dependent manner. Phosphoghrelin and phosphodesacylghrelin show greatly diminished phosphatidylserine-dependent binding. These results are consistent with binding of ghrelin and desacylghrelin to acidic lipids via the basic face of an amphipathic helix with Ser-18 phosphorylation disrupting both helical character and membrane binding.

KW - Ghrelin phosphorylation

KW - Ghrelin structure

KW - PKC substrate

KW - Peptide-membrane binding

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JO - Peptides

JF - Peptides

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ER -

Regulation of ghrelin structure and membrane binding by phosphorylation

Abstract

Keywords

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