Isolation, characterization and molecular cloning of new temporins from the skin of the North African ranid Pelophylax saharica.
Temporins are small antimicrobial peptides isolated from North American and Eurasian ranid frogs that are particularly active against Gram-positive bacteria. To date, no temporins have been characterized from North African frog species. We isolated three novel members of the temporin family, named temporin-1Sa (FLSGIVGMLGKLF(amide)), -1Sb (FLPIVTNLLSGLL(amide)), and -1Sc (FLSHIAGFLSNLF(amide)), from the skin of the Sahara frog Pelophylax (Rana) saharica originating from Tunisia. These temporins were identified by a combined mass spectrometry/molecular cloning approach. Temporin-1Sa was found to be highly active against Gram-positive and Gram-negative bacteria, yeasts and fungi (MIC=2-30 microM). To our knowledge, this is the first 13-residue member of the temporin family with a net charge of +2 that shows such broad-spectrum activity with particularly high potency on the clinically relevant Gram-negative strains, Escherichia coli (MIC=10 microM) and Pseudomonas aeruginosa (MIC=31 microM). Moreover, temporin-1Sa displays significant antiparasitic activity (IC50 approximately 20 microM) against the promastigote and amastigote stages of Leishmania infantum.
Abbassi
Feten
F
FRE 2852, Peptidome de la Peau des Amphibiens, Université Pierre et Marie Curie/CNRS, Tour 43, 2 place Jussieu, 75251 Paris Cedex 05, France.
Oury
Bruno
B
Blasco
Thierry
T
Sereno
Denis
D
Bolbach
Gérard
G
Nicolas
Pierre
P
Hani
Khaled
K
Amiche
Mohamed
M
Ladram
Ali
A
D000595:Amino Acid Sequence; D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D000935:Antifungal Agents / Q000494:pharmacology; D023181:Antimicrobial Cationic Peptides / Q000302:isolation & purification / Q000494:pharmacology; D000981:Antiprotozoal Agents / Q000494:pharmacology; D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D004912:Erythrocytes / Q000187:drug effects; D053878:Hemolytic Agents / Q000494:pharmacology; D006801:Humans; D018314:Leishmania infantum / Q000187:drug effects; D008969:Molecular Sequence Data; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification*; D011898:Ranidae
D000900:Anti-Bacterial Agents; D000935:Antifungal Agents; D023181:Antimicrobial Cationic Peptides; D000981:Antiprotozoal Agents; D053878:Hemolytic Agents; D011506:Proteins; C104501:temporin