Identification of Components in Frog Skin Secretions with Therapeutic Potential as Antidiabetic Agents

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Abstract

Several peptides that were first identified on the basis of their antimicrobial or immunomodulatory properties have subsequently shown potential for development into agents for the treatment of patients with Type 2 diabetes. A strategy is presented for the isolation and characterization of such peptides in norepinephrine- stimulated skin secretions from a range of frog species. The methodology involves fractionation of the secretions by reversed-phase HPLC, identification of fractions containing components that stimulate the rate of release of insulin from BRIN-BD11 clonal β-cells without simultaneously stimulating the release of lactate dehydrogenase, identification of active peptides in the mass range 1–6 kDa by MALDI-TOF mass spectrometry, purification of the peptides to near homogeneity by further HPLC, and structural characterization by automated Edman degradation. The effect of synthetic replicates of the active peptides on glucose homeostasis in vivo may be evaluated in mice fed a high fat diet to produce obesity, glucose intolerance, and insulin resistance.
LanguageEnglish
Pages319-333
JournalMethods in Molecular Biology
Volume1719
Early online date24 Feb 2018
DOIs
Publication statusE-pub ahead of print - 24 Feb 2018

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Hypoglycemic Agents
Skin
Peptides
Insulin
Glucose
Fractionation
Nutrition
Medical problems
L-Lactate Dehydrogenase
Purification
Mass spectrometry
Norepinephrine
Fats
Degradation

Keywords

  • Frog skin secretions
  • Antidiabetic peptide
  • BRIN-BD11 cells
  • Insulin
  • Incretin

Cite this

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title = "Identification of Components in Frog Skin Secretions with Therapeutic Potential as Antidiabetic Agents",
abstract = "Several peptides that were first identified on the basis of their antimicrobial or immunomodulatory properties have subsequently shown potential for development into agents for the treatment of patients with Type 2 diabetes. A strategy is presented for the isolation and characterization of such peptides in norepinephrine- stimulated skin secretions from a range of frog species. The methodology involves fractionation of the secretions by reversed-phase HPLC, identification of fractions containing components that stimulate the rate of release of insulin from BRIN-BD11 clonal β-cells without simultaneously stimulating the release of lactate dehydrogenase, identification of active peptides in the mass range 1–6 kDa by MALDI-TOF mass spectrometry, purification of the peptides to near homogeneity by further HPLC, and structural characterization by automated Edman degradation. The effect of synthetic replicates of the active peptides on glucose homeostasis in vivo may be evaluated in mice fed a high fat diet to produce obesity, glucose intolerance, and insulin resistance.",
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author = "JM Conlon and Charlotte Moffett and J Leprince and Peter Flatt",
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AU - Moffett, Charlotte

AU - Leprince, J

AU - Flatt, Peter

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AB - Several peptides that were first identified on the basis of their antimicrobial or immunomodulatory properties have subsequently shown potential for development into agents for the treatment of patients with Type 2 diabetes. A strategy is presented for the isolation and characterization of such peptides in norepinephrine- stimulated skin secretions from a range of frog species. The methodology involves fractionation of the secretions by reversed-phase HPLC, identification of fractions containing components that stimulate the rate of release of insulin from BRIN-BD11 clonal β-cells without simultaneously stimulating the release of lactate dehydrogenase, identification of active peptides in the mass range 1–6 kDa by MALDI-TOF mass spectrometry, purification of the peptides to near homogeneity by further HPLC, and structural characterization by automated Edman degradation. The effect of synthetic replicates of the active peptides on glucose homeostasis in vivo may be evaluated in mice fed a high fat diet to produce obesity, glucose intolerance, and insulin resistance.

KW - Frog skin secretions

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KW - Incretin

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