Antimicrobial peptide defenses of the Tarahumara frog, Rana tarahumarae.
Populations of the Tarahumara frog Rana tarahumarae have decreased markedly in recent years in the northern part of their range. Infection by the chytrid fungus Batrachochytrium dendrobatidis has been implicated in these declines. To determine whether antimicrobial peptides in the skin provide protection against this pathogen, norepinephrine-stimulated skin secretions were tested for their ability to inhibit growth of B. dendrobatidis in vitro. After concentration, crude mixtures of skin peptides inhibited the growth of the chytrid in a concentration-dependent manner. Proteomic analysis led to the identification and characterization of three peptides belonging to the brevinin-1 family of antimicrobial peptides and three belonging to the ranatuerin-2 family. The two most abundant peptides, ranatuerin-2TRa (GIMDSIKGAAKEIAGHLLDNLKCKITGC) and brevinin-1TRa (FLPVIAGIAANVLPKLFCKLTKRC), were active against B. dendrobatidis (MIC of 50 microM for ranatuerin-2TRa and 12.5 microM for brevinin-1TRa against zoospores). These data clearly show that antimicrobial peptides in the skin secretions of the Tarahumara frog are active against B. dendrobatidis and should provide some protection against infection. Therefore, the observed susceptibility of these frogs to this pathogen in the wild may be due to the effects of additional environmental factors that impair this innate defense mechanism, leading to the observed population declines.
Rollins-Smith
Louise A
LA
Department of Microbiology, A-5301 Medical Center North, Vanderbilt University, Nashville, TN 37232, USA. louise.rollins-smith@vanderbilt.edu
Reinert
Laura K
LK
Miera
Verma
V
Conlon
J Michael
JM
D000595:Amino Acid Sequence; D029845:Amphibian Proteins; D000818:Animals; D000935:Antifungal Agents / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D023181:Antimicrobial Cationic Peptides / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D005658:Fungi / Q000187:drug effects* / Q000472:pathogenicity; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D010455:Peptides; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D011898:Ranidae / Q000378:metabolism* / Q000382:microbiology; D012867:Skin / Q000737:chemistry*
D029845:Amphibian Proteins; D000935:Antifungal Agents; D023181:Antimicrobial Cationic Peptides; D010455:Peptides; D011506:Proteins; C121230:ranatuerin; C078413:brevinin-1 protein, Rana