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Frontiers in endocrinology2024; 15; 1411483; doi: 10.3389/fendo.2024.1411483

Acyl modifications in bovine, porcine, and equine ghrelins.

Abstract: Ghrelin is a peptide hormone with various important physiological functions. The unique feature of ghrelin is its serine 3 acyl-modification, which is essential for ghrelin activity. The major form of ghrelin is modified with n-octanoic acid (C8:0) by ghrelin O-acyltransferase. Various acyl modifications have been reported in different species. However, the underlying mechanism by which ghrelin is modified with various fatty acids remains to be elucidated. Herein, we report the purification of bovine, porcine, and equine ghrelins. The major active form of bovine ghrelin was a 27-amino acid peptide with an n-octanoyl (C8:0) modification at Ser3. The major active form of porcine and equine ghrelin was a 28-amino acid peptide. However, porcine ghrelin was modified with n-octanol (C8:0), whereas equine ghrelin was modified with n-butanol (C4:0) at Ser3. This study indicates the existence of structural divergence in ghrelin and suggests that it is necessary to measure the minor and major forms of ghrelin to fully understand its physiology.
Copyright © 2024 Ida, Tominaga, Iwamoto, Kurogi, Okura, Shimada, Kato, Kuwano, Ode, Nagata, Kitamura, Yazawa, Sato-Hashimoto, Yasuda, Miyazato, Shiimura, Sato and Kojima.
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Publication Date: 2024-05-17 PubMed ID: 38828411PubMed Central: PMC11140078DOI: 10.3389/fendo.2024.1411483Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

Objective Overview

  • This study examined the specific fatty acid modifications on the peptide hormone ghrelin from cattle, pigs, and horses, revealing differences in the type and length of acyl groups attached, which are important for the hormone’s activity and may impact our understanding of its physiological roles.

Introduction to Ghrelin and Acyl Modification

  • Ghrelin is a peptide hormone involved in various crucial physiological processes, such as regulating appetite, energy balance, and growth hormone secretion.
  • A distinctive feature of ghrelin is the acyl modification at the serine residue in the third position (Ser3), which is critical for the hormone’s biological activity.
  • This acylation typically involves the attachment of a fatty acid, most commonly n-octanoic acid (C8:0), catalyzed by the enzyme ghrelin O-acyltransferase (GOAT).
  • Previous research has identified various acyl modifications of ghrelin in different species, but the mechanisms governing the type of fatty acid attached were unclear.

Research Aim and Methods

  • The study aimed to purify ghrelin from three species: bovine (cattle), porcine (pig), and equine (horse), and analyze the acyl modifications present.
  • Purification processes were carried out to isolate the active forms of ghrelin peptides from each species.
  • The peptide length and the specific acyl groups attached at Ser3 were determined, likely using biochemical techniques such as mass spectrometry and peptide sequencing.

Key Findings

  • Bovine Ghrelin:
    • The major active form consisted of a 27-amino acid peptide.
    • The peptide had an n-octanoyl (C8:0) modification at Ser3, consistent with the common modification seen in many species.
  • Porcine Ghrelin:
    • The major active form was a 28-amino acid peptide.
    • The acyl modification at Ser3 was also n-octanoyl (C8:0), matching the bovine modification in fatty acid type.
  • Equine Ghrelin:
    • The major active form was likewise a 28-amino acid peptide.
    • The acyl modification differed, composed of n-butanoyl (C4:0) at Ser3, which is a shorter chain fatty acid compared to the octanoyl group.

Implications and Conclusions

  • The data reveal structural divergence in ghrelin across species, particularly in the fatty acid modifications attached at the Ser3 site.
  • The porcine and bovine ghrelins share the common octanoyl (C8:0) modification despite a difference in peptide length, whereas equine ghrelin differs in both peptide length and the nature of the acyl modification (shorter chain, C4:0).
  • This divergence suggests species-specific differences in ghrelin’s biosynthesis, possibly reflecting variations in the enzyme substrate specificity or availability of fatty acid substrates.
  • Measuring both the major and minor forms of ghrelin, including different acyl variants, is necessary to fully understand the physiological actions and regulatory mechanisms of this hormone in various species.
  • Understanding the diversity in acyl modifications may impact future research on ghrelin function, potential diagnostic measures, and therapeutic approaches in veterinary and human medicine.

Cite This Article

APA
Ida T, Tominaga H, Iwamoto E, Kurogi A, Okura A, Shimada K, Kato J, Kuwano A, Ode H, Nagata S, Kitamura K, Yazawa T, Sato-Hashimoto M, Yasuda M, Miyazato M, Shiimura Y, Sato T, Kojima M. (2024). Acyl modifications in bovine, porcine, and equine ghrelins. Front Endocrinol (Lausanne), 15, 1411483. https://doi.org/10.3389/fendo.2024.1411483

Publication

Frontiers in endocrinology
ISSN: 1664-2392
NlmUniqueID: 101555782
Country: Switzerland
Language: English
Volume: 15
Pages: 1411483
PII: 1411483

Researcher Affiliations

Ida, Takanori
  • Division for Identification and Analysis of Bioactive Peptides, Department of Bioactive Peptides, Frontier Science Research Center, University of Miyazaki, Miyazaki, Japan.
Tominaga, Hatsumi
  • Miyazaki Prefecture Industrial Technology Center, Miyazaki, Japan.
Iwamoto, Eri
  • Clinical Research Center, Kurume University Hospital, Fukuoka, Japan.
Kurogi, Akito
  • Division for Identification and Analysis of Bioactive Peptides, Department of Bioactive Peptides, Frontier Science Research Center, University of Miyazaki, Miyazaki, Japan.
Okura, Ayaka
  • Division for Identification and Analysis of Bioactive Peptides, Department of Bioactive Peptides, Frontier Science Research Center, University of Miyazaki, Miyazaki, Japan.
Shimada, Kengo
  • Division for Identification and Analysis of Bioactive Peptides, Department of Bioactive Peptides, Frontier Science Research Center, University of Miyazaki, Miyazaki, Japan.
Kato, Johji
  • Division for Identification and Analysis of Bioactive Peptides, Department of Bioactive Peptides, Frontier Science Research Center, University of Miyazaki, Miyazaki, Japan.
Kuwano, Atsutoshi
  • Equine Research Institute, Japan Racing Association, Tochigi, Japan.
Ode, Hirotaka
  • Racehorse Clinic, Ritto Training Center, Japan Racing Association, Shiga, Japan.
Nagata, Sayaka
  • Department of Food Science and Technology, Faculty of Health and Nutrition, Minami Kyushu University, Miyazaki, Japan.
Kitamura, Kazuo
  • Department of Projects Research, Frontier Science Research Center, University of Miyazaki, Miyazaki, Japan.
Yazawa, Takashi
  • Department of Biochemistry, Asahikawa Medical University, Hokkaido, Japan.
Sato-Hashimoto, Miho
  • Department of Animal Pharmaceutical Science, School of Pharmaceutical Sciences, Kyusyu University of Medical Science, Miyazaki, Japan.
Yasuda, Masahiro
  • Laboratory of Veterinary Anatomy, Faculty of Agriculture, University of Miyazaki, Miyazaki, Japan.
Miyazato, Mikiya
  • Division for Identification and Analysis of Bioactive Peptides, Department of Bioactive Peptides, Frontier Science Research Center, University of Miyazaki, Miyazaki, Japan.
Shiimura, Yuki
  • Molecular Genetics, Institute of Life Sciences, Kurume University, Fukuoka, Japan.
Sato, Takahiro
  • Molecular Genetics, Institute of Life Sciences, Kurume University, Fukuoka, Japan.
Kojima, Masayasu
  • Molecular Genetics, Institute of Life Sciences, Kurume University, Fukuoka, Japan.

MeSH Terms

  • Animals
  • Ghrelin / metabolism
  • Ghrelin / chemistry
  • Horses
  • Cattle
  • Swine
  • Amino Acid Sequence
  • Acylation
  • Caprylates / metabolism

Conflict of Interest Statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

References

This article includes 40 references
  1. Kojima M, Hosoda H, Date Y, Nakazato M, Matsuo H, Kangawa K. Ghrelin is a growth-hormone-releasing acylated peptide from stomach.. Nature (1999) 402:656–60.
    doi: 10.1038/45230pubmed: 10604470google scholar: lookup
  2. Angeloni SV, Glynn N, Ambrosini G, Garant MJ, Higley JD, Suomi S. Characterization of the rhesus monkey ghrelin gene and factors influencing ghrelin gene expression and fasting plasma levels.. Endocrinology (2004) 145:2197–205.
    doi: 10.1210/en.2003-1103pubmed: 14736731google scholar: lookup
  3. Galas L, Chartrel N, Kojima M, Kangawa K, Vaudry H. Immunohistochemical localization and biochemical characterization of ghrelin in the brain and stomach of the frog Rana esculenta.. J Comp Neurol (2002) 450:34–44.
    doi: 10.1002/cne.10291pubmed: 12124765google scholar: lookup
  4. Kaiya H, Kojima M, Hosoda H, Moriyama S, Takahashi A, Kawauchi H. Peptide purification, complementary deoxyribonucleic acid (DNA) and genomic DNA cloning, and functional characterization of ghrelin in rainbow trout.. Endocrinology (2003) 144:5215–26.
    doi: 10.1210/en.2003-1085pubmed: 12970156google scholar: lookup
  5. Unniappan S, Lin X, Cervini L, Rivier J, Kaiya H, Kangawa K. Goldfish ghrelin: Molecular characterization of the complementary deoxyribonucleic acid, partial gene structure and evidence for its stimulatory role in food intake.. Endocrinology (2002) 143:4143–6.
    doi: 10.1210/en.2002-220644pubmed: 12239128google scholar: lookup
  6. Kaiya H, Kojima M, Hosoda H, Riley LG, Hirano T, Grau EG. Identification of tilapia ghrelin and its effects on growth hormone and prolactin release in the tilapia, Oreochromis mossambicus.. Comp Biochem Physiol B Biochem Mol Biol (2003) 135:421–9.
    doi: 10.1016/S1096-4959(03)00109-Xpubmed: 12831762google scholar: lookup
  7. Kaiya H, Kojima M, Hosoda H, Riley LG, Hirano T, Grau EG. Amidated fish ghrelin: Purification, cdna cloning in the Japanese eel and its biological activity.. J Endocrinol (2003) 176:415–23.
    doi: 10.1677/joe.0.1760415pubmed: 12630926google scholar: lookup
  8. Parhar IS, Sato H, Sakuma Y. Ghrelin gene in cichlid fish is modulated by sex and development.. Biochem Biophys Res Commun (2003) 305:169–75.
    doi: 10.1016/S0006-291X(03)00729-0pubmed: 12732213google scholar: lookup
  9. Tanaka M, Hayashida Y, Iguchi T, Nakao N, Nakai N, Nakashima K. Organization of the mouse ghrelin gene and promoter: Occurrence of a short noncoding first exon.. Endocrinology (2001) 142:3697–700.
    doi: 10.1210/endo.142.8.8433pubmed: 11459820google scholar: lookup
  10. Yabuki A, Ojima T, Kojima M, Nishi Y, Mifune H, Matsumoto M. Characterization and species differences in gastric ghrelin cells from mice, rats and hamsters.. J Anat (2004) 205:239–46.
    doi: 10.1111/j.0021-8782.2004.00331.xpmc: PMC1571341pubmed: 15379929google scholar: lookup
  11. Palyha OC, Feighner SD, Tan CP, McKee KK, Hreniuk DL, Gao YD. Ligand activation domain of human orphan growth hormone (GH) secretagogue receptor (GHS-R) conserved from pufferfish to humans.. Mol Endocrinol (2000) 14:160–9.
    doi: 10.1210/mend.14.1.0412pubmed: 10628755google scholar: lookup
  12. Chan CB, Cheng CHK. Identification and functional characterization of two alternatively spliced growth hormone secretagogue receptor transcripts from the pituitary of black seabream Acanthopagrus schlegeli.. Mol Cell Endocrinol (2004) 214:81–95.
    doi: 10.1016/j.mce.2003.11.020pubmed: 15062547google scholar: lookup
  13. Tanaka M, Miyazaki T, Yamamoto I, Nakai N, Ohta Y, Tsushima N. Molecular characterization of chicken growth hormone secretagogue receptor gene.. Gen Comp Endocrinol (2003) 134:198–202.
    doi: 10.1016/S0016-6480(03)00247-8pubmed: 14511992google scholar: lookup
  14. Gnanapavan S, Kola B, Bustin SA, Morris DG, McGee P, Fairclough P. The tissue distribution of the mRNA of ghrelin and subtypes of its receptor, GHS-R, in humans.. J Clin Endocrinol Metab (2002) 87:2988.
    doi: 10.1210/jcem.87.6.8739pubmed: 12050285google scholar: lookup
  15. Guan XM, Yu H, Palyha OC, McKee KK, Feighner SD, Sirinathsinghji DJS. Distribution of mrna encoding the growth hormone secretagogue receptor in brain and peripheral tissues.. Brain Res Mol Brain Res (1997) 48:23–9.
    doi: 10.1016/S0169-328X(97)00071-5pubmed: 9379845google scholar: lookup
  16. Bennett PA, Thomas GB, Howard AD, Feighner SD, van der Ploeg LHT, Smith RG. Hypothalamic growth hormone secretagogue-receptor (GHS-R) expression is regulated by growth hormone in the rat.. Endocrinology (1997) 138:4552–7.
    doi: 10.1210/endo.138.11.5476pubmed: 9348177google scholar: lookup
  17. Nakazato M, Murakami N, Date Y, Kojima M, Matsuo H, Kangawa K. A role for ghrelin in the central regulation of feeding.. Nature (2001) 409:194–8.
    doi: 10.1038/35051587pubmed: 11196643google scholar: lookup
  18. Sato T, Oishi K, Koga D, Ida T, Sakai Y, Kangawa K. Thermoregulatory role of ghrelin in the induction of torpor under a restricted feeding condition.. Sci Rep (2021) 11:17954.
    doi: 10.1038/s41598-021-97440-ypmc: PMC8438062pubmed: 34518616google scholar: lookup
  19. Masuda Y, Tanaka T, Inomata N, Ohnuma N, Tanaka S, Itoh Z. Ghrelin stimulates gastric acid secretion and motility in rats.. Biochem Biophys Res Commun (2000) 276:905–8.
    doi: 10.1006/bbrc.2000.3568pubmed: 11027567google scholar: lookup
  20. Date Y, Nakazato M, Murakami N, Kojima M, Kangawa K, Matsukura S. Ghrelin acts in the central nervous system to stimulate gastric acid secretion.. Biochem Biophys Res Commun (2001) 280:904–7.
    doi: 10.1006/bbrc.2000.4212pubmed: 11162609google scholar: lookup
  21. Lee HM, Wang G, Englander EW, Kojima M, Greeley GH. Ghrelin, a new gastrointestinal endocrine peptide that stimulates insulin secretion: Enteric distribution, ontogeny, influence of endocrine, and dietary manipulations.. Endocrinology (2002) 143:185–90.
    doi: 10.1210/endo.143.1.8602pubmed: 11751608google scholar: lookup
  22. Nagaya N, Kojima M, Uematsu M, Yamagishi M, Hosoda H, Oya H. Hemodynamic and hormonal effects of human ghrelin in healthy volunteers.. Am J Physiol Regul Integr Comp Physiol (2001) 280:R1483–7.
    doi: 10.1152/ajpregu.2001.280.5.R1483pubmed: 11294772google scholar: lookup
  23. Nagaya N, Uematsu M, Kojima M, Ikeda Y, Yoshihara F, Shimizu W. Chronic administration of ghrelin improves left ventricular dysfunction and attenuates development of cardiac cachexia in rats with heart failure.. Circulation (2001) 104:1430–5.
    doi: 10.1161/hc3601.095575pubmed: 11560861google scholar: lookup
  24. Nakahara K, Nakagawa M, Baba Y, Sato M, Toshinai K, Date Y. Maternal ghrelin plays an important role in rat fetal development during pregnancy.. Endocrinology (2006) 147:1333–42.
    doi: 10.1210/en.2005-0708pubmed: 16339208google scholar: lookup
  25. Yang J, Brown MS, Liang G, Grishin NV, Goldstein JL. Identification of the acyltransferase that Octanoylates ghrelin, an appetite-stimulating peptide hormone.. Cell (2008) 132:387–96.
    doi: 10.1016/j.cell.2008.01.017pubmed: 18267071google scholar: lookup
  26. Ohgusu H, Shirouzu K, Nakamura Y, Nakashima Y, Ida T, Sato T. Ghrelin O-acyltransferase (GOAT) has a preference for n-hexanoyl-CoA over n-octanoyl-CoA as an acyl donor.. Biochem Biophys Res Commun (2009) 386:153–8.
    doi: 10.1016/j.bbrc.2009.06.001pubmed: 19501572google scholar: lookup
  27. Kaiya H, Kojima M, Hosoda H, Koda A, Yamamoto K, Kitajima Y. Bullfrog ghrelin is modified by n-octanoic acid at its third threonine residue.. J Biol Chem (2001) 276:40441–8.
    doi: 10.1074/jbc.M105212200pubmed: 11546772google scholar: lookup
  28. Kaiya H, van der Geyten S, Kojima M, Hosoda H, Kitajima Y, Matsumoto M. Chicken ghrelin: Purification, cDNA cloning, and biological activity.. Endocrinology (2002) 143:3454–63.
    doi: 10.1210/en.2002-220255pubmed: 12193558google scholar: lookup
  29. Matsumoto M, Hosoda H, Kitajima Y, Morozumi N, Minamitake Y, Tanaka S. Structure-activity relationship of ghrelin: Pharmacological study of ghrelin peptides.. Biochem Biophys Res Commun (2001) 287:142–6.
    doi: 10.1006/bbrc.2001.5553pubmed: 11549267google scholar: lookup
  30. Hosoda H, Kojima M, Matsuo H, Kangawa K. Purification and characterization of rat des-Gln14-ghrelin, a second endogenous ligand for the growth hormone secretagogue receptor.. J Biol Chem (2000) 275:21995–2000.
    doi: 10.1074/jbc.M002784200pubmed: 10801861google scholar: lookup
  31. Hosoda H, Kojima M, Mizushima T, Shimizu S, Kangawa K. Structural divergence of human ghrelin: Identification of multiple ghrelin-derived molecules produced by post-translational processing.. J Biol Chem (2003) 278:64–70.
    doi: 10.1074/jbc.M205366200pubmed: 12414809google scholar: lookup
  32. Ida T, Miyazato M, Naganobu K, Nakahara K, Sato M, Lin XZ. Purification and characterization of feline ghrelin and its possible role.. Domest Anim Endocrinol (2007) 32:93–105.
    doi: 10.1016/j.domaniend.2006.01.002pubmed: 16466902google scholar: lookup
  33. Ida T, Miyazato M, Lin XZ, Kaiya H, Sato T, Nakahara K. Purification and characterization of caprine ghrelin and its effect on growth hormone release.. J Mol Neurosci (2010) 42:99–105.
    doi: 10.1007/s12031-010-9379-0pubmed: 20437258google scholar: lookup
  34. Ishida Y, Sakahara S, Tsutsui C, Kaiya H, Sakata I, Oda Si. Identification of ghrelin in the house musk shrew (Suncus murinus): cDNA cloning, peptide purification and tissue distribution.. Pept (N Y) (2009) 30:982–90.
    doi: 10.1016/j.peptides.2009.01.006pubmed: 19428777google scholar: lookup
  35. Liu H, Sun D, Myasnikov A, Damian M, Baneres JL, Sun J. Structural basis of human ghrelin receptor signaling by ghrelin and the synthetic agonist ibutamoren.. Nat Commun (2021) 12:6410.
    doi: 10.1038/s41467-021-26735-5pmc: PMC8568970pubmed: 34737341google scholar: lookup
  36. Wang Y, Guo S, Zhuang Y, Yun Y, Xu P, He X. Molecular recognition of an acyl-peptide hormone and activation of ghrelin receptor.. Nat Commun (2021) 12:5064.
    doi: 10.1038/s41467-021-25364-2pmc: PMC8379176pubmed: 34417468google scholar: lookup
  37. Shiimura Y, Horita S, Hamamoto A, Asada H, Hirata K, Tanaka M. Structure of an antagonist-bound ghrelin receptor reveals possible ghrelin recognition mode.. Nat Commun (2020) 11:4160.
    doi: 10.1038/s41467-020-17554-1pmc: PMC7438500pubmed: 32814772google scholar: lookup
  38. Nishi Y, Hiejima H, Hosoda H, Kaiya H, Mori K, Fukue Y. Ingested medium-chain fatty acids are directly utilized for the acyl modification of ghrelin.. Endocrinology (2005) 146:2255–64.
    doi: 10.1210/en.2004-0695pubmed: 15677766google scholar: lookup
  39. Hanis F, Chung ELT, Kamalludin MH, Idrus Z. Blood profile, hormones, and telomere responses: Potential biomarkers in horses exhibiting abnormal oral behavior.. J Equine Vet Sci (2022) 118:104130.
    doi: 10.1016/j.jevs.2022.104130pubmed: 36182046google scholar: lookup
  40. Moore JL, Siciliano PD, Pratt-Phillips SE. Effects of diet versus exercise on morphometric measurements, blood hormone concentrations, and oral sugar test response in obese horses.. J Equine Vet Sci (2019) 78:38–45.
    doi: 10.1016/j.jevs.2019.03.214pubmed: 31203982google scholar: lookup

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