Peptidomic analysis of skin secretions from the bullfrog Lithobates catesbeianus (Ranidae) identifies multiple peptides with potent insulin-releasing activity

Milena Mechkarska, Opeolu O. Ojo, Mohammed A. Meetani, Laurent Coquet, Thierry Jouenne, Yasser Abdel-Wahab, Peter Flatt, Jay D. King, J. Michael Conlon

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

Using a combination of reversed-phase HPLC and electrospray mass spectrometry, peptidomic analysis of norepinephrine-stimulated skin secretions of the American bullfrog Litho bates catesbeianus Shaw, 1802 led to the identification and characterization of five newly described peptides (ranatuerin-1CBb, ranatuerin-2CBc, and -CBd, palustrin-2CBa, and temporin-CBf) together with seven peptides previously isolated on the basis of their antimicrobial activity (ranatuerin-1CBa, ranatuerin-2CBa, brevinin-1CBa, and -1CBb, temporin-CBa, -CBb, and -CBd). The abilities of the most abundant of the purified peptides to stimulate the release of insulin from the rat BRIN-BD11 clonal beta cell line were evaluated. Ranatuerin-2CBd (GFLDIIKNLGKTFAGHMLDKIRCTIGTCPPSP) was the most potent peptide producing a significant stimulation of insulin release (119% of basal rate, P<0.01) from BRIN-BD11 cells at a concentration of 30 nM, with a maximum response (236% of basal rate, P<0.001) at a concentration of 3 mu M. Ranatuerin-2CBd did not stimulate release of the cytosolic enzyme, lactate dehydrogenase at concentrations up to 3 mu M, indicating that the integrity of the plasma membrane had been preserved. Brevinin-1CBb (FLPFIAR-LAAKVFPSIICSVTKKC) produced the maximum stimulation of insulin release (285% of basal rate, P<0.001 at 3 mu M) but the peptide was cytotoxic at this concentration. (C) 2010 Elsevier Inc. All rights reserved.
LanguageEnglish
Pages203-208
JournalPeptides
Volume32
Issue number2
DOIs
Publication statusPublished - Feb 2011

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Ranidae
Lithobates catesbeianus
insulin
secretion
peptides
reversed-phase high performance liquid chromatography
norepinephrine
lactate dehydrogenase
plasma membrane
anti-infective agents
cell lines
mass spectrometry
rats
enzymes
cells

Cite this

Mechkarska, Milena ; Ojo, Opeolu O. ; Meetani, Mohammed A. ; Coquet, Laurent ; Jouenne, Thierry ; Abdel-Wahab, Yasser ; Flatt, Peter ; King, Jay D. ; Conlon, J. Michael. / Peptidomic analysis of skin secretions from the bullfrog Lithobates catesbeianus (Ranidae) identifies multiple peptides with potent insulin-releasing activity. In: Peptides. 2011 ; Vol. 32, No. 2. pp. 203-208.
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abstract = "Using a combination of reversed-phase HPLC and electrospray mass spectrometry, peptidomic analysis of norepinephrine-stimulated skin secretions of the American bullfrog Litho bates catesbeianus Shaw, 1802 led to the identification and characterization of five newly described peptides (ranatuerin-1CBb, ranatuerin-2CBc, and -CBd, palustrin-2CBa, and temporin-CBf) together with seven peptides previously isolated on the basis of their antimicrobial activity (ranatuerin-1CBa, ranatuerin-2CBa, brevinin-1CBa, and -1CBb, temporin-CBa, -CBb, and -CBd). The abilities of the most abundant of the purified peptides to stimulate the release of insulin from the rat BRIN-BD11 clonal beta cell line were evaluated. Ranatuerin-2CBd (GFLDIIKNLGKTFAGHMLDKIRCTIGTCPPSP) was the most potent peptide producing a significant stimulation of insulin release (119{\%} of basal rate, P<0.01) from BRIN-BD11 cells at a concentration of 30 nM, with a maximum response (236{\%} of basal rate, P<0.001) at a concentration of 3 mu M. Ranatuerin-2CBd did not stimulate release of the cytosolic enzyme, lactate dehydrogenase at concentrations up to 3 mu M, indicating that the integrity of the plasma membrane had been preserved. Brevinin-1CBb (FLPFIAR-LAAKVFPSIICSVTKKC) produced the maximum stimulation of insulin release (285{\%} of basal rate, P<0.001 at 3 mu M) but the peptide was cytotoxic at this concentration. (C) 2010 Elsevier Inc. All rights reserved.",
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Peptidomic analysis of skin secretions from the bullfrog Lithobates catesbeianus (Ranidae) identifies multiple peptides with potent insulin-releasing activity. / Mechkarska, Milena; Ojo, Opeolu O.; Meetani, Mohammed A.; Coquet, Laurent; Jouenne, Thierry; Abdel-Wahab, Yasser; Flatt, Peter; King, Jay D.; Conlon, J. Michael.

In: Peptides, Vol. 32, No. 2, 02.2011, p. 203-208.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Peptidomic analysis of skin secretions from the bullfrog Lithobates catesbeianus (Ranidae) identifies multiple peptides with potent insulin-releasing activity

AU - Mechkarska, Milena

AU - Ojo, Opeolu O.

AU - Meetani, Mohammed A.

AU - Coquet, Laurent

AU - Jouenne, Thierry

AU - Abdel-Wahab, Yasser

AU - Flatt, Peter

AU - King, Jay D.

AU - Conlon, J. Michael

PY - 2011/2

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N2 - Using a combination of reversed-phase HPLC and electrospray mass spectrometry, peptidomic analysis of norepinephrine-stimulated skin secretions of the American bullfrog Litho bates catesbeianus Shaw, 1802 led to the identification and characterization of five newly described peptides (ranatuerin-1CBb, ranatuerin-2CBc, and -CBd, palustrin-2CBa, and temporin-CBf) together with seven peptides previously isolated on the basis of their antimicrobial activity (ranatuerin-1CBa, ranatuerin-2CBa, brevinin-1CBa, and -1CBb, temporin-CBa, -CBb, and -CBd). The abilities of the most abundant of the purified peptides to stimulate the release of insulin from the rat BRIN-BD11 clonal beta cell line were evaluated. Ranatuerin-2CBd (GFLDIIKNLGKTFAGHMLDKIRCTIGTCPPSP) was the most potent peptide producing a significant stimulation of insulin release (119% of basal rate, P<0.01) from BRIN-BD11 cells at a concentration of 30 nM, with a maximum response (236% of basal rate, P<0.001) at a concentration of 3 mu M. Ranatuerin-2CBd did not stimulate release of the cytosolic enzyme, lactate dehydrogenase at concentrations up to 3 mu M, indicating that the integrity of the plasma membrane had been preserved. Brevinin-1CBb (FLPFIAR-LAAKVFPSIICSVTKKC) produced the maximum stimulation of insulin release (285% of basal rate, P<0.001 at 3 mu M) but the peptide was cytotoxic at this concentration. (C) 2010 Elsevier Inc. All rights reserved.

AB - Using a combination of reversed-phase HPLC and electrospray mass spectrometry, peptidomic analysis of norepinephrine-stimulated skin secretions of the American bullfrog Litho bates catesbeianus Shaw, 1802 led to the identification and characterization of five newly described peptides (ranatuerin-1CBb, ranatuerin-2CBc, and -CBd, palustrin-2CBa, and temporin-CBf) together with seven peptides previously isolated on the basis of their antimicrobial activity (ranatuerin-1CBa, ranatuerin-2CBa, brevinin-1CBa, and -1CBb, temporin-CBa, -CBb, and -CBd). The abilities of the most abundant of the purified peptides to stimulate the release of insulin from the rat BRIN-BD11 clonal beta cell line were evaluated. Ranatuerin-2CBd (GFLDIIKNLGKTFAGHMLDKIRCTIGTCPPSP) was the most potent peptide producing a significant stimulation of insulin release (119% of basal rate, P<0.01) from BRIN-BD11 cells at a concentration of 30 nM, with a maximum response (236% of basal rate, P<0.001) at a concentration of 3 mu M. Ranatuerin-2CBd did not stimulate release of the cytosolic enzyme, lactate dehydrogenase at concentrations up to 3 mu M, indicating that the integrity of the plasma membrane had been preserved. Brevinin-1CBb (FLPFIAR-LAAKVFPSIICSVTKKC) produced the maximum stimulation of insulin release (285% of basal rate, P<0.001 at 3 mu M) but the peptide was cytotoxic at this concentration. (C) 2010 Elsevier Inc. All rights reserved.

U2 - 10.1016/j.peptides.2010.11.002

DO - 10.1016/j.peptides.2010.11.002

M3 - Article

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SP - 203

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JO - Peptides

T2 - Peptides

JF - Peptides

SN - 0196-9781

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