TY - JOUR
T1 - Purification and characterization of antimicrobial peptides from the skin secretions of the carpenter frog Rana virgatipes (Ranidae, Aquarana)
AU - Conlon, J. Michael
AU - Abraham, Bency
AU - Sonnevend, Agnes
AU - Jouenne, Thierry
AU - Cosette, Pascal
AU - Leprince, Jerome
AU - Vaudry, Hubert
AU - Bevier, Catherine R.
N1 - Funding Information:
This work was supported by an Interdisciplinary Grant (03/12-8-03-01) and a Faculty Support Grant (NP/05/01) from the United Arab Emirates University to JMC and a grant from the Natural Science Division at Colby College and funds from the Clare Boothe Luce Program of the Henry Luce Foundation to CRB. The authors thank Laurey Steinke and Michele Fontaine (University of Nebraska Medical Center, Omaha, NE) for amino acid composition analysis and Mac Given (Neumann College), Hande Bartuoglu, Dan Breen, and Lauren Henderson (Colby College) for help with secretion collection in the field.
PY - 2005/11/15
Y1 - 2005/11/15
N2 - The members of the Aquarana (or Rana catesbeiana species group) form a well-supported monophyletic clade but phylogenetic relationships between species within the group are incompletely understood. Peptides that differentially inhibited the growth of bacteria were purified from electrically stimulated skin secretions of the carpenter frog Rana virgatipes. Structural characterization identified members of the ranatuerin-2 (3 peptides) and temporin (3-peptides) families, previously found in the skins of R. catesbeiana, R. clamitans, R. grylio and R. septentrionalis. Ranalexin, a peptide previously found only in the Aquarana, was isolated together with a variant (FFGLHNLVPSMLCVVRKKC) that lacks the propensity to adopt an α-helical conformation and so was devoid of antimicrobial activity. Two C-terminally α-amidated peptides belonging to the brevinin-2 family were isolated from the skin secretions that, like an ortholog from R. septentrionalis, lacked the C-terminal cyclic heptapeptide domain associated with members of this family. Ranatuerin-1, previously isolated from R. catesbeiana, R. clamitans and R. grylio but absent from R. septentrionalis, was also not identified in R. virgatipes. Synthetic replicates of temporin-1Va (FLSSIGKILGNLL.NH2), temporin-IVb (FLSIIAKVLGSLF.NH2) and temporin-1Vc (FLPLVTMLLGKLF.NH2) potently inhibited growth of Gram-positive bacteria (including methicillin-resistant Staphylococcus aureus). Temporin-1Va was also active against Gram-negative bacteria and the opportunistic yeast pathogen Candida albicans and had relatively weak hemolytic activity (LD50 = 120 μM) and may therefore represent a candidate for drug development. Our data support the placement of R. virgatipes in the Aquarana and indicate a closer phylogenetic relationship of R. virgatipes with R. septentrionalis than with R. catesbeiana, R. clamitans and R. grylio.
AB - The members of the Aquarana (or Rana catesbeiana species group) form a well-supported monophyletic clade but phylogenetic relationships between species within the group are incompletely understood. Peptides that differentially inhibited the growth of bacteria were purified from electrically stimulated skin secretions of the carpenter frog Rana virgatipes. Structural characterization identified members of the ranatuerin-2 (3 peptides) and temporin (3-peptides) families, previously found in the skins of R. catesbeiana, R. clamitans, R. grylio and R. septentrionalis. Ranalexin, a peptide previously found only in the Aquarana, was isolated together with a variant (FFGLHNLVPSMLCVVRKKC) that lacks the propensity to adopt an α-helical conformation and so was devoid of antimicrobial activity. Two C-terminally α-amidated peptides belonging to the brevinin-2 family were isolated from the skin secretions that, like an ortholog from R. septentrionalis, lacked the C-terminal cyclic heptapeptide domain associated with members of this family. Ranatuerin-1, previously isolated from R. catesbeiana, R. clamitans and R. grylio but absent from R. septentrionalis, was also not identified in R. virgatipes. Synthetic replicates of temporin-1Va (FLSSIGKILGNLL.NH2), temporin-IVb (FLSIIAKVLGSLF.NH2) and temporin-1Vc (FLPLVTMLLGKLF.NH2) potently inhibited growth of Gram-positive bacteria (including methicillin-resistant Staphylococcus aureus). Temporin-1Va was also active against Gram-negative bacteria and the opportunistic yeast pathogen Candida albicans and had relatively weak hemolytic activity (LD50 = 120 μM) and may therefore represent a candidate for drug development. Our data support the placement of R. virgatipes in the Aquarana and indicate a closer phylogenetic relationship of R. virgatipes with R. septentrionalis than with R. catesbeiana, R. clamitans and R. grylio.
KW - Antimicrobial peptide
KW - Brevinin-2
KW - Frog skin
KW - Ranalexin
KW - Ranatuerin-2
KW - Temporin
UR - http://www.scopus.com/inward/record.url?scp=25844481594&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=25844481594&partnerID=8YFLogxK
U2 - 10.1016/j.regpep.2005.06.003
DO - 10.1016/j.regpep.2005.06.003
M3 - Article
C2 - 15996769
AN - SCOPUS:25844481594
SN - 0167-0115
VL - 131
SP - 38
EP - 45
JO - Regulatory Peptides
JF - Regulatory Peptides
IS - 1-3
ER -