1. Primary structures of skin antimicrobial peptides indicate a close, but not conspecific, phylogenetic relationship between the leopard frogs Lithobates onca and Lithobates yavapaiensis (Ranidae)
J Michael Conlon, Laurent Coquet, Jérôme Leprince, Thierry Jouenne, Hubert Vaudry, Jay D King Comp Biochem Physiol C Toxicol Pharmacol. 2010 Apr;151(3):313-7. doi: 10.1016/j.cbpc.2009.12.004. Epub 2009 Dec 31.
The phylogenetic relationship between the relict leopard frog Lithobates (Rana) onca (Cope, 1875) and the lowland leopard frog Lithobates (Rana) yavapaiensis (Platz and Frost, 1984) is unclear. Chromatographic analysis of norepinephrine-stimulated skin secretions from L. onca led to the identification of six peptides with antimicrobial activity. Determination of their primary structures indicated that four of the peptides were identical to brevinin-1Ya, brevinin-1Yb, brevinin-1Yc and ranatuerin-2Ya previously isolated from skin secretions of L. yavapaiensis. However, a peptide belonging to the temporin family (temporin-ONa: FLPTFGKILSGLF.NH(2)) and an atypical member of the ranatuerin-2 family containing a C-terminal cyclic heptapeptide domain (ranatuerin-2ONa: GLMDTVKNAAKNLAGQMLDKLKCKITGSC) were isolated from the L. onca secretions but were not present in the L. yavapaiensis secretions. Ranatuerin-2ONa inhibited the growth of Escherichia coli (MIC=50muM) and Candida albicans (MIC=100muM ) and showed hemolytic activity (LC(50)=90muM) but was inactive against Staphylococcus aureus. The data indicate a close phylogenetic relationship between L. onca and L. yavapaiensis but suggest that they are not conspecific species.
2. Peptides with potent cytolytic activity from the skin secretions of the North American leopard frogs, Lithobates blairi and Lithobates yavapaiensis
J Michael Conlon, Eman Ahmed, Laurent Coquet, Thierry Jouenne, Jérôme Leprince, Hubert Vaudry, Jay D King Toxicon. 2009 Jun;53(7-8):699-705. doi: 10.1016/j.toxicon.2009.02.018. Epub 2009 Feb 28.
Six structurally similar and strongly cationic peptides belonging to the brevinin-1 family were isolated from skin secretions of the plains leopard frog Lithobates blairi and the lowland leopard frog Lithobates yavapaiensis on the basis of their antimicrobial activities. Brevinin-1BLc (FLPIIAGIAAKFLPKIFCTISKKC) from L. blairi represented the most potent peptide (MIC=25microM Escherichia coli, MIC=1.5microM Staphylococcus aureus, MIC=3microM Candida albicans, LC(50)=9microM human erythrocytes and LC(50)=6microM HepG2 human hepatoma-derived cells). The appreciably lower antimicrobial potencies of brevinin-1Ya and -1Yc from L. yavapaiensis correlate with the decreases in cationicity produced by the amino acid substitutions Lys(11)-->Asn (brevinin-1Ya) and Pro(14)-->Glu (brevinin-1Yc). In addition, a peptide isolated from the skin secretions of L. yavapaiensis belonging to the ranatuerin-2 family (ranatuerin-2Ya; GLMDTIKGVAKTVAASWLDKLKCKIT GC) inhibited the growth of E. coli (MIC=50microM) and S. aureus (MIC=50microM). In contrast to brevinin-1BLc, ranatuerin-2Ya showed appreciably greater cytolytic activity against HepG2 cells (LC(50)=20microM) than against erythrocytes (LC(50)>100microM).
