title: Identification and characterization of antimicrobial peptides from skin of Amolops ricketti (Anura: Ranidae).
abstract: As one of large amphibian group, there are a total of 45 species of Amolops in the world. However, the antimicrobial peptides (AMPs) existing in this genus has not been extensively studied. In this study, cDNAs encoding five novel AMP precursors were cloned by screening the skin-derived cDNA library of Amolops ricketti, a frog species that exists in southern and western parts of China. Protein sequence analysis led to the identification of five deduced peptides, three belonging to the brevinin-1 family and two belonging to the brevinin-2 family of amphibian AMPs. Thus, they were named as brevinin-1RTa (FLPLLAGVVANFLPQIICKIARKC), brevinin-1RTb (FLGSLLGLVGKVVPTLFCKISKKC), brevinin-1RTc (FLGSLLGLVGKIVPTLICKISKKC), brevinin-2RTa (GLMSTLKDFGKTAAKEIAQSLLSTASCKLAKTC), and brevinin-2RTb (GILDTLKEFGKTAAKGIAQSLLSTASCKLAKTC), respectively. The purification of brevinin-1RTa, brevinin-1RTb, and brevinin-2RTb was carried out by RP-HPLC, and confirmed by the LC-MS/MS-based proteomics approach. All of the peptides displayed different antimicrobial potency against a variety of microorganisms. In addition, brevinin-2RTa and brevinin-2RTb were found to have relatively low hemolytic activity (>400μg/ml) against mammalian red blood cells in vitro, which could potentially be as candidates for developing novel anti-infection agents.
authors.affiliation: Key Laboratory of Animal Physiology, Biochemistry and Molecular Biology of Hebei Province, College of Life Sciences, Hebei Normal University, Shijiazhuang, Hebei 050016, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Wang
authors.forename: Ran
authors.initials: R
authors.lastname: Ran
authors.forename: Haining
authors.initials: H
authors.lastname: Yu
authors.forename: Zhijun
authors.initials: Z
authors.lastname: Yu
authors.forename: Yuhong
authors.initials: Y
authors.lastname: Hu
authors.forename: Hongyuan
authors.initials: H
authors.lastname: Zheng
authors.forename: Duo
authors.initials: D
authors.lastname: Wang
authors.forename: Fan
authors.initials: F
authors.lastname: Yang
authors.forename: Renjie
authors.initials: R
authors.lastname: Liu
authors.forename: Jingze
authors.initials: J
authors.lastname: Liu
mesh_terms: D000595:Amino Acid Sequence; D029845:Amphibian Proteins / Q000737:chemistry / Q000235:genetics; D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000737:chemistry* / Q000235:genetics* / Q000494:pharmacology*; D001483:Base Sequence; D002176:Candida albicans / Q000187:drug effects; D003001:Cloning, Molecular; D018076:DNA, Complementary; D005260:Female; D053878:Hemolytic Agents / Q000737:chemistry / Q000494:pharmacology; D008297:Male; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D017433:Protein Structure, Secondary; D011898:Ranidae / Q000235:genetics*; D012867:Skin / Q000737:chemistry; D013211:Staphylococcus aureus / Q000187:drug effects; D053719:Tandem Mass Spectrometry
chemical_list: D029845:Amphibian Proteins; D023181:Antimicrobial Cationic Peptides; D018076:DNA, Complementary; D053878:Hemolytic Agents; C078413:brevinin-1 protein, Rana; C078414:brevinin-2, Rana