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
Y1 - 2011/2
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
SN - 1873-5169
VL - 32
SP - 203
EP - 208
JO - Peptides
JF - Peptides
IS - 2
ER -