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Molecular Cloning of a Novel Tryptophyllin Peptide from the Skin of the Orange‐Legged Monkey Frog, Phyllomedusa hypochondrialis
Author(s) -
Wang Ran,
Lin Yangjun,
Chen Tianbao,
zhou Mei,
Wang Lei,
Shaw Chris
Publication year - 2014
Publication title -
chemical biology and drug design
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.59
H-Index - 77
eISSN - 1747-0285
pISSN - 1747-0277
DOI - 10.1111/cbdd.12287
Subject(s) - complementary dna , edman degradation , peptide , peptide sequence , microbiology and biotechnology , amino acid , biology , biochemistry , cdna library , frog skin , chemistry , gene , organic chemistry , sodium
Tryptophyllins are a group of small (4–14 amino acids), heterogenous peptides, mostly from the skins of hylid frogs from the genera, Phyllomedusa and Litoria . To date, more than forty TPHs have been discovered in species from these two genera. Here, we describe the identification of a novel tryptophyllin type 3 peptide, PhT‐3, from the extracts of skin of the orange‐legged monkey frog, Phyllomedusa hypochondrialis , and molecular cloning of its precursor‐encoding cDNA from a cDNA library constructed from the same skin sample. Full primary structural characterization was achieved using a combination of direct Edman degradation, mass spectrometry and deduction from cloned skin‐derived cDNA . The open‐reading frame of the precursor cDNA was found to consist of 63 amino acid residues. The mature peptide arising from this precursor contains a post‐translationally modified N‐terminal pyroglutamate ( pG lu) residue, formed from acid‐mediated cyclization of an N‐terminal Gln (Q) residue, and with the structure: pG lu‐Asp‐Lys‐Pro‐Phe‐Trp‐Pro‐Pro‐Pro‐Ile‐Tyr‐Pro‐Met. Pharmacological assessment of a synthetic replicate of this peptide on phenylephrine preconstricted rat tail artery segments, revealed a reduction in relaxation induced by bradykinin. PhT‐3 was also found to mediate antiproliferative effects on human prostate cancer cell lines.