Antimicrobial peptides from the skin of the Japanese mountain brown frog Rana ornativentris: Evidence for polymorphism among preprotemporin mRNAs

A previous study led to the isolation of antimicrobial peptides belonging to the temporin and brevinin-2 families from a pooled extract of the skin of adult specimens of the Japanese mountain brown frog Rana ornativentris Werner 1903. In order to ascertain whether individual frogs expressed the full...

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Bibliographic Details
Published in:Peptides (New York, N.Y. : 1980) Vol. 28; no. 3; pp. 524 - 532
Main Authors: Ohnuma, Aya, Michael Conlon, J., Yamaguchi, Keiko, Kawasaki, Hiroaki, Coquet, Laurent, Leprince, Jérôme, Jouenne, Thierry, Vaudry, Hubert, Iwamuro, Shawichi
Format: Journal Article
Language:English
Published: United States Elsevier Inc 01.03.2007
Elsevier
Subjects:
Amino Acid Sequence
Animals
Antimicrobial Cationic Peptides
Antimicrobial Cationic Peptides - genetics
Antimicrobial Cationic Peptides - isolation & purification
Antimicrobial Cationic Peptides - pharmacology
Antimicrobial peptide
Base Sequence
Cell Behavior
Cellular Biology
Chemical Sciences
Cloning, Molecular
DNA, Complementary
DNA, Complementary - genetics
Female
Frog skin
Gene polymorphism
Japan
Life Sciences
Male
Medicinal Chemistry
Microbial Sensitivity Tests
Molecular Sequence Data
Neurobiology
Neurons and Cognition
Pharmaceutical sciences
Pharmacology
Polymorphism, Genetic
Preprotemporin
Protein Precursors
Protein Precursors - genetics
Protein Precursors - isolation & purification
Proteins
Proteins - genetics
Proteins - isolation & purification
Proteins - pharmacology
Ranidae
Ranidae - genetics
Ranidae - metabolism
RNA, Messenger
RNA, Messenger - genetics
Sequence Homology, Amino Acid
Sequence Homology, Nucleic Acid
Skin
Skin - metabolism
Japan
Frog skin
Gene polymorphism
Antimicrobial peptide
Preprotemporin
ISSN:0196-9781, 1873-5169
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Summary:A previous study led to the isolation of antimicrobial peptides belonging to the temporin and brevinin-2 families from a pooled extract of the skin of adult specimens of the Japanese mountain brown frog Rana ornativentris Werner 1903. In order to ascertain whether individual frogs expressed the full complement of temporin genes, we individually cloned cDNAs encoding the temporin precursors from total RNA extracted from the skins of 12 frogs by RT-PCR using a set of preprotemporin-specific primers. All the specimens examined contained mRNAs directing the synthesis of the novel, but inactive, temporin-1Oe (ILPLLGNLLNGLL·NH 2). Nucleotide sequence analysis revealed marked polymorphism among individual frogs. Twenty-seven distinct preprotemporin-1Oe mRNAs were identified that contained synonymous substitutions in the antimicrobial peptide region and both synonymous and non-synonymous substitutions in the signal peptide and intervening sequence regions. Up to eight preprotemporin-1Oe mRNA variants were found within a single frog. In addition, several cDNAs encoding preprotemporin-1Oa and -1Ob and a single cDNA encoding preprotemporin-1Oc were characterized. Peptidomic analysis of norepinephrine-stimulated skin secretions revealed the presence of temporin-1Oe, temporin-1Of (SLILKGLASIAKLF·NH 2), temporin-1Og (FLSSLLSKVVSLFT·NH 2), four members of the ranatuerin-2 family and one member of the palustrin-2 family in addition to previously characterized temporin and brevinin-2 peptides.
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ISSN:0196-9781
1873-5169
DOI:10.1016/j.peptides.2006.10.013