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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Vydané v:	Peptides (New York, N.Y. : 1980) Ročník 28; číslo 3; s. 524 - 532
Hlavní autori:	Ohnuma, Aya, Michael Conlon, J., Yamaguchi, Keiko, Kawasaki, Hiroaki, Coquet, Laurent, Leprince, Jérôme, Jouenne, Thierry, Vaudry, Hubert, Iwamuro, Shawichi
Médium:	Journal Article
Jazyk:	English
Vydavateľské údaje:	United States Elsevier Inc 01.03.2007 Elsevier
Predmet:	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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Abstract	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.
AbstractList	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 x NH2). 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 x NH2), temporin-1Og (FLSSLLSKVVSLFT x NH2), 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. 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 x NH2). 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 x NH2), temporin-1Og (FLSSLLSKVVSLFT x NH2), 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.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 x NH2). 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 x NH2), temporin-1Og (FLSSLLSKVVSLFT x NH2), 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. 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.
Author	Iwamuro, Shawichi Ohnuma, Aya Kawasaki, Hiroaki Michael Conlon, J. Vaudry, Hubert Jouenne, Thierry Coquet, Laurent Leprince, Jérôme Yamaguchi, Keiko
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Keywords	Frog skin Gene polymorphism Antimicrobial peptide Preprotemporin
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Snippet	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...
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SubjectTerms	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
Title	Antimicrobial peptides from the skin of the Japanese mountain brown frog Rana ornativentris: Evidence for polymorphism among preprotemporin mRNAs
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