Advertisement

Amino Acids

, Volume 50, Issue 6, pp 723–734 | Cite as

Insulinotropic, glucose-lowering, and beta-cell anti-apoptotic actions of peptides related to esculentin-1a(1-21).NH2

  • Vishal Musale
  • Yasser H. A. Abdel-Wahab
  • Peter R. Flatt
  • J. Michael ConlonEmail author
  • Maria Luisa Mangoni
Original Article

Abstract

Long-standing Type 2 diabetes is associated with loss of both β‐cell function and β‐cell mass. Peptides derived from the frog-skin host-defense peptide esculentin-1 have been shown to exhibit potent, broad-spectrum antimicrobial activity. The aim of the present study is to determine whether such peptides also show insulinotropic and β-cell protective activities. Esculentin-1a(1-21).NH2, esculentin-1b(1-18).NH2, and esculentin-1a(1-14).NH2 produced concentration-dependent stimulations of insulin release from BRIN-BD11 rat clonal β-cells, 1.1B4 human-derived pancreatic β-cells, and isolated mouse islets with no cytotoxicity at concentrations of up to 3 μM. The mechanism of insulinotropic action involved membrane depolarization and an increase in intracellular Ca2+ concentrations. The analogue [D-Lys14, D-Ser17]esculentin-1a(1-21).NH2 (Esc(1-21)-1c) was less potent in vitro than the all L-amino acid containing peptides and esculentin-1a(9-21) was inactive indicating that helicity is an important determinant of insulinotropic activity. However, intraperitoneal injection of Esc(1-21)-1c (75 nmol/kg body weight) together with a glucose load (18 mmol/kg body weight) in C57BL6 mice improved glucose tolerance with a concomitant increase in insulin secretion, whereas administration of esculentin-1a(1-21).NH2, esculentin-1b(1-18).NH2, and esculentin-1a(1-14) was without significant effect on plasma glucose levels. Esc(1-21)-1c (1 µM) protected BRIN-BD11 cells against cytokine-induced apoptosis (P < 0.01) and augmented proliferation of the cells (P < 0.01) to a similar extent as glucagon-like peptide-1. The data demonstrate that the multifunctional peptide Esc(1-21)-1c, as well as showing therapeutic potential as an anti-infective and wound-healing agent, may constitute a template for development of compounds for treatment of patients with Type 2 diabetes.

Keywords

Esculentin-1a(1-21) Type 2 diabetes Amphibian skin peptide Insulin release β-cell proliferation Anti-apoptotic peptide 

Notes

Acknowledgements

Funding for this study was provided by a project Grant from Diabetes UK (12/0004457) and by the University of Ulster Research Strategy Funding.

Compliance with ethical standards

Ethical approval

This article does not contain any studies with human participants performed by any of the authors. For this type of study, informed consent is not relevant. All animal experiments were carried out in accordance with the UK Animals (Scientific Procedures) Act 1986 and EU Directive 2010/63EU for animal experiments and approved by Ulster University Animal Ethics Review Committee. All necessary steps were taken to prevent any potential animal suffering.

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

726_2018_2551_MOESM1_ESM.tif (670 kb)
Supplementary material 1 (TIFF 670 kb) Supplementary Fig. S1. Effects of Esc(1-21)-1c (1 µM) and KCl (30 mM) on (A) membrane potential in BRIN-BD11 cells expressed as relative fluorescence units, RFU and (B) the integrated response (area under the curve, AUC). Values are mean ± SEM (n = 6). *** P < 0.001 compared with 5.6 mM glucose alone
726_2018_2551_MOESM2_ESM.tif (696 kb)
Supplementary material 2 (TIFF 696 kb) Supplementary Fig. S2. Effects of Esc(1-21)-1c (1 µM) and alanine (10 mM) on (A) intracellular calcium ion concentrations [Ca2+]i in BRIN-BD11 cells expressed as relative fluorescence units, RFU and (B) the integrated response (area under the curve, AUC). Values are mean ± SEM (n = 6). ***P < 0.001 compared with 5.6 mM glucose alone

References

  1. Arden C (2018) A role for glucagon-like peptide-1 in the regulation of β-cell autophagy. Peptides 100:85–93CrossRefPubMedGoogle Scholar
  2. Cappiello F, Di Grazia A, Segev-Zarko LA, Scali S, Ferrera L, Galietta L, Pini A, Shai Y, Di YP, Mangoni ML (2016) Esculentin-1a-derived peptides promote clearance of Pseudomonas aeruginosa internalized in bronchial cells of cystic fibrosis patients and lung cell migration: biochemical properties and a plausible mode of action. Antimicrob Agents Chemother 60:7252–7262PubMedPubMedCentralGoogle Scholar
  3. Casciaro B, Dutta D, Loffredo MR, Marcheggiani S, McDermott AM, Willcox MD, Mangoni ML (2018) Esculentin-1a derived peptides kill Pseudomonas aeruginosa biofilm on soft contact lenses and retain antibacterial activity upon immobilization to the lens surface. Biopolymers (in press) Google Scholar
  4. Chen C, Mangoni ML, Di YP (2017) In vivo therapeutic efficacy of frog skin-derived peptides against Pseudomonas aeruginosa-induced pulmonary infection. Sci Rep 7:8548CrossRefPubMedPubMedCentralGoogle Scholar
  5. Conlon JM, Kolodziejek J, Nowotny N (2009) Antimicrobial peptides from the skins of North American frogs. Biochim Biophys Acta 1788:1556–1563CrossRefPubMedGoogle Scholar
  6. Conlon JM, Mechkarska M, Lukic ML, Flatt PR (2014) Potential therapeutic applications of multifunctional host-defense peptides from frog skin as anti-cancer, anti-viral, immunomodulatory, and anti-diabetic agents. Peptides 57:67–77CrossRefPubMedGoogle Scholar
  7. Conlon JM, Mechkarska M, Abdel-Wahab YH, Flatt PR (2018) Peptides from frog skin with potential for development into agents for Type 2 diabetes therapy. Peptides 100:275–281CrossRefPubMedGoogle Scholar
  8. Di Grazia A, Cappiello F, Cohen H, Casciaro B, Luca V, Pini A, Di YP, Shai Y, Mangoni ML (2015a) D-Amino acids incorporation in the frog skin-derived peptide esculentin-1a(1-21)NH2 is beneficial for its multiple functions. Amino Acids 47:2505–2519CrossRefPubMedGoogle Scholar
  9. Di Grazia A, Cappiello F, Imanishi A, Mastrofrancesco A, Picardo M, Paus R, Mangoni ML (2015b) The frog skin-derived antimicrobial peptide esculentin-1a(1-21)NH2 promotes the migration of human HaCaT keratinocytes in an EGF receptor-dependent manner: a novel promoter of human skin wound healing? PLoS One 10:e0128663CrossRefPubMedPubMedCentralGoogle Scholar
  10. Flatt PR, Bailey CJ (1981) Abnormal plasma glucose and insulin responses in heterozygous lean (ob/+) mice. Diabetologia 20:573–577CrossRefPubMedGoogle Scholar
  11. Ghosh A, Bera S, Shai Y, Mangoni ML, Bhunia A (2016) NMR structure and binding of esculentin-1a (1-21)NH2 and its diastereomer to lipopolysaccharide: correlation with biological functions. Biochim Biophys Acta 1858:800–812CrossRefPubMedGoogle Scholar
  12. Goto M, Maki T, Kiyoizumi T, Satomi S, Monaco AP (1985) An improved method for isolation of mouse pancreatic islets. Transplantation 40:437–438CrossRefGoogle Scholar
  13. Graham GV, Conlon JM, Abdel-Wahab YH, Gault VA, Flatt PR (2018) Evaluation of the insulinotropic and glucose-lowering actions of zebrafish GIP in mammalian systems: evidence for involvement of the GLP-1 receptor. Peptides 100:182–189CrossRefPubMedGoogle Scholar
  14. Graz H, D’Souza VK, Alderson DEC, Graz M (2017) Diabetes-related amputations create considerable public health burden in the UK. Diabetes Res Clin Pract 135:158–165CrossRefPubMedGoogle Scholar
  15. Islas-Rodrìguez AE, Marcellini L, Orioni B, Barra D, Stella L, Mangoni ML (2009) Esculentin 1-21: a linear antimicrobial peptide from frog skin with inhibitory effect on bovine mastitis-causing bacteria. J Pept Sci 15:607–614CrossRefPubMedGoogle Scholar
  16. Khan D, Vasu S, Moffett RC, Irwin N, Flatt PR (2016) Islet distribution of Peptide YY and its regulatory role in primary mouse islets and immortalized rodent and human beta-cell function and survival. Mol Cell Endocrinol 436:102–113CrossRefPubMedGoogle Scholar
  17. Kolar SS, Luca V, Baidouri H, Mannino G, McDermott AM, Mangoni ML (2015) Esculentin-1a(1-21)NH2: a frog skin-derived peptide for microbial keratitis. Cell Mol Life Sci 72:617–627CrossRefPubMedGoogle Scholar
  18. Kyte J, Doolittle DF (1982) A simple method for displaying the hydropathic character of a protein. J Mol Biol 157:105–132CrossRefPubMedGoogle Scholar
  19. Lee YS, Jun HS (2014) Anti-diabetic actions of glucagon-like peptide-1 on pancreatic beta-cells. Metabolism 63:9–19CrossRefPubMedGoogle Scholar
  20. Loffredo MR, Ghosh A, Harmouche N, Casciaro B, Luca V, Bortolotti A, Cappiello F, Stella L, Bhunia A, Bechinger B, Mangoni ML (2017) Membrane perturbing activities and structural properties of the frog-skin derived peptide esculentin-1a(1-21)NH2 and its diastereomer Esc(1-21)-1c: correlation with their antipseudomonal and cytotoxic activity. Biochim Biophys Acta 1859:2327–2339CrossRefPubMedGoogle Scholar
  21. Luca V, Stringaro A, Colone M, Pini A, Ml Mangoni (2013) Esculentin(1-21), an amphibian skin membrane-active peptide with potent activity on both planktonic and biofilm cells of the bacterial pathogen Pseudomonas aeruginosa. Cell Mol Life Sci 70:2773–2778CrossRefPubMedGoogle Scholar
  22. Luca V, Olivi M, Di Grazia A, Palleschi C, Uccelletti D, Mangoni ML (2014) Anti-Candida activity of 1-18 fragment of the frog skin peptide esculentin-1b: in vitro and in vivo studies in a Caenorhabditis elegans infection model. Cell Mol Life Sci 71:2535–2546PubMedGoogle Scholar
  23. Maisetta G, Mangoni ML, Esin S, Pichierri G, Capria AL, Brancatisano FL, Di Luca M, Barnini S, Barra D, Campa M, Batoni G (2009) In vitro bactericidal activity of the N-terminal fragment of the frog peptide esculentin-1b (Esc 1-18) in combination with conventional antibiotics against Stenotrophomonas maltophilia. Peptides 30:1622–1626CrossRefPubMedGoogle Scholar
  24. Mangoni ML, Fiocco D, Mignogna G, Barra D, Simmaco M (2003) Functional characterisation of the 1-18 fragment of esculentin-1b, an antimicrobial peptide from Rana esculenta. Peptides 24:1771–1777CrossRefPubMedGoogle Scholar
  25. Mangoni ML, Luca V, McDermott AM (2015) Fighting microbial infections: a lesson from amphibian skin-derived esculentin-1 peptides. Peptides 71:286–295CrossRefPubMedGoogle Scholar
  26. Manzo G, Casu M, Rinaldi AC, Montaldo NP, Luganini A, Gribaudo G, Scorciapino MA (2014) Folded structure and insertion depth of the frog-skin antimicrobial peptide esculentin-1b(1-18) in the presence of differently charged membrane-mimicking micelles. J Nat Prod 77:2410–2417CrossRefPubMedGoogle Scholar
  27. Marcellini L, Borro M, Gentile G, Rinaldi AC, Stella L, Aimola P, Barra D, Mangoni ML (2009) Esculentin-1b(1-18)–a membrane-active antimicrobial peptide that synergizes with antibiotics and modifies the expression level of a limited number of proteins in Escherichia coli. FEBS J 276:5647–5664CrossRefPubMedGoogle Scholar
  28. Marenah L, Flatt PR, Orr DF, Shaw C, Abdel-Wahab YH (2006) Skin secretions of Rana saharica frogs reveal antimicrobial peptides esculentins-1 and -1B and brevinins-1E and -2EC with novel insulin releasing activity. J Endocrinol 188:1–9CrossRefPubMedGoogle Scholar
  29. McClenaghan NH, Barnett CR, Ah-Sing E, Abdel-Wahab YHA, O’Harte FP, Yoon TW, Swanston-Flatt SK, Flatt PR (1996) Characterization of a novel glucose-responsive insulin-secreting cell line, BRIN-BD11, produced by electrofusion. Diabetes 45:1132–1140CrossRefPubMedGoogle Scholar
  30. McCluskey JT, Hamid M, Guo-Parke H, McClenaghan NH, Gomis R, Flatt PR (2011) Development and functional characterization of insulin-releasing human pancreatic beta cell lines produced by electrofusion. J Biol Chem 286:21982–21992CrossRefPubMedPubMedCentralGoogle Scholar
  31. Miguel JC, Patterson S, Abdel-Wahab YH, Mathias PC, Flatt PR (2004) Time-correlation between membrane depolarization and intracellular calcium in insulin secreting BRIN-BD11 cells: studies using FLIPR. Cell Calcium 36:43–50CrossRefPubMedGoogle Scholar
  32. Muller LM, Gorter KJ, Hak E, Goudzwaard WL, Schellevis FG, Hoepelman AI, Rutten GE (2005) Increased risk of common infections in patients with Type 1 and Type 2 diabetes mellitus. Clin Infect Dis 41:281–288CrossRefPubMedGoogle Scholar
  33. Muñoz V, Serrano L (1994) Elucidating the folding problem of helical peptides using empirical parameters. Nature Struct Biol 1:399–409CrossRefPubMedGoogle Scholar
  34. Ojo OO, Srinivasan DK, Owolabi BO, McGahon MK, Moffett RC, Curtis TM, Conlon JM, Flatt PR, Abdel-Wahab YH (2016) Molecular mechanisms mediating the beneficial metabolic effects of [Arg4]tigerinin-1R in mice with diet-induced obesity and insulin resistance. Biol Chem 397:753–764CrossRefPubMedGoogle Scholar
  35. Owolabi BO, Musale V, Ojo OO, Moffett RC, McGahon MK, Curtis TM, Conlon JM, Flatt PR, Abdel-Wahab YHA (2017) Actions of PGLa-AM1 and its [A14 K] and [A20 K] analogues and their therapeutic potential as anti-diabetic agents. Biochimie 138:1–12CrossRefPubMedGoogle Scholar
  36. Pantic JM, Jovanovic IP, Radosavljevic GD, Arsenijevic NN, Conlon JM, Lukic ML (2017) The potential of frog skin-derived peptides for development into therapeutically-valuable immunomodulatory agents. Molecules 22:E2071CrossRefPubMedGoogle Scholar
  37. Ponti D, Mignogna G, Mangoni ML, De Biase D, Simmaco M, Barra D (1999) Expression and activity of cyclic and linear analogues of esculentin-1, an anti-microbial peptide from amphibian skin. Eur J Biochem 263:921–927CrossRefPubMedGoogle Scholar
  38. Ponti D, Mangoni ML, Mignogna G, Simmaco M, Barra D (2003) An amphibian antimicrobial peptide variant expressed in Nicotiana tabacum confers resistance to phytopathogens. Biochem J 370:121–127CrossRefPubMedPubMedCentralGoogle Scholar
  39. Ríos JL, Francini F, Schinella GR (2015) Natural products for the treatment of type 2 diabetes mellitus. Planta Med 81:975–994CrossRefPubMedGoogle Scholar
  40. Rollins-Smith LA, Carey C, Longcore J, Doersam JK, Boutte A, Bruzgal JE, Conlon JM (2002) Activity of antimicrobial skin peptides from ranid frogs against Batrachochytrium dendrobatidis, the chytrid fungus associated with global amphibian declines. Dev Comp Immunol 26:471–479CrossRefPubMedGoogle Scholar
  41. Simmaco M, Mignogna G, Barra D, Bossa F (1994) Antimicrobial peptides from skin secretions of Rana esculenta. Molecular cloning of cDNAs encoding esculentin and brevinins and isolation of new active peptides. J Biol Chem 269:11956–11961PubMedGoogle Scholar
  42. Srinivasan D, Ojo OO, Abdel-Wahab YH, Flatt PR, Guilhaudis L, Conlon JM (2014) Insulin-releasing and cytotoxic properties of the frog skin peptide, tigerinin-1R: a structure-activity study. Peptides 55:23–31CrossRefPubMedGoogle Scholar
  43. Sun J, Xu M, Ortsäter H, Lundeberg E, Juntti-Berggren L, Chen YQ, Haeggström JZ, Gudmundsson GH, Diana J, Agerberth B (2016) Cathelicidins positively regulate pancreatic β-cell functions. FASEB J 30:884–894CrossRefPubMedGoogle Scholar
  44. Vasu S, McGahon MK, Moffett RC, Curtis TM, Conlon JM, Abdel-Wahab YH, Flatt PR (2017a) Esculentin-2CHa(1-30) and its analogues: stability and mechanisms of insulinotropic action. J Endocrinol 232:423–435CrossRefPubMedGoogle Scholar
  45. Vasu S, Ojo OO, Moffett RC, Conlon JM, Flatt PR, Abdel-Wahab YHA (2017b) Anti-diabetic actions of esculentin-2CHa(1-30) and its stable analogues in a diet-induced model of obesity-diabetes. Amino Acids 49:1705–1717CrossRefPubMedGoogle Scholar
  46. Xu X, Lai R (2015) The chemistry and biological activities of peptides from amphibian skin secretions. Chem Rev 115:1760–1846CrossRefPubMedGoogle Scholar
  47. Yabe D, Seino Y (2011) Two incretin hormones GLP-1 and GIP: comparison of their actions in insulin secretion and β cell preservation. Prog Biophys Mol Biol 107:248–256CrossRefPubMedGoogle Scholar
  48. Zummo FP, Cullen KS, Honkanen-Scott M, Shaw JAM, Lovat PE, Arden C (2017) Glucagon-like peptide 1 protects pancreatic β-cells from death by increasing autophagic flux and restoring lysosomal function. Diabetes 66:1272–1285CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2018

Authors and Affiliations

  • Vishal Musale
    • 1
  • Yasser H. A. Abdel-Wahab
    • 1
  • Peter R. Flatt
    • 1
  • J. Michael Conlon
    • 1
    Email author
  • Maria Luisa Mangoni
    • 2
  1. 1.SAAD Centre for Pharmacy and Diabetes, School of Biomedical SciencesUniversity of UlsterColeraineUK
  2. 2.Laboratory Affiliated To Istituto Pasteur Italia-Fondazione Cenci Bolognetti, Department of Biochemical SciencesSapienza University of RomeRomeItaly

Personalised recommendations