Molecular cloning and sequence analyses of preprotemporin mRNAs containing premature stop codons from extradermal tissues of Rana tagoi

Shawichi Iwamuro, Marico Nakamura, Aya Ohnuma, J. Michael Conlon

Research output: Contribution to journalArticlepeer-review

22 Citations (Scopus)

Abstract

The temporins are a family of hydrophobic, C-terminally α-amidated antimicrobial peptides that are synthesized in the skins of a wide range of species of frogs belonging to the genus Rana. In the present study, we investigated using RT-PCR the expression of preprotemporin mRNAs in extradermal tissues of Tago's brown frog Rana tagoi. cDNAs encoding temporin-1TGa (FLPILGKLLS10GIL.NH2), previously isolated from an extract of the skin of R. tagoi skin, were amplified and cloned from the stomach, liver, kidney, skeletal muscle. However, a net insertion of 10 nucleotides resulted in the presence of a premature stop codon in the open reading frame that was not present in the corresponding region of preprotemporin-1TGa from skin. The preprotemporin cDNA obtained from small intestine contained an additional 12 nucleotide insertion in the region that encodes the temporin sequence so that a novel peptide (FLPVILPVIG10KLLSGIL.NH2), termed temporin-1TGc, is specified. This cDNA also contained a premature stop codon in the open reading frame. Although it is unclear whether temporin-1TGc is produced in R. tagoi tissues, a synthetic replicate of the peptide of was biologically active, inhibiting the growth of Staphylococcus aureus (minimal inhibitory concentration = 37.5 μM) and producing hemolysis of human erythrocytes (LD50 = 50 μM).

Original languageEnglish
Pages (from-to)2124-2128
Number of pages5
JournalPeptides
Volume27
Issue number9
DOIs
Publication statusPublished - Sep 2006
Externally publishedYes

Keywords

  • Antimicrobial peptide
  • Rana tagoi
  • Temporin

ASJC Scopus subject areas

  • Biochemistry
  • Physiology
  • Endocrinology
  • Cellular and Molecular Neuroscience

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