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Title: The chains of the heterodimeric amphibian skin antimicrobial peptide, distinctin, are encoded by separate messenger RNAs
Authors: Evaristo, Geisa
Pinkse, Martijn
Wang, Lei
Zhou, Mei
Wu, Youjia
Wang, Hui
Chen, Tianbao
Shaw, Chris
Issue Date: 2013
Source: JOURNAL OF PROTEOMICS, 78, p. 245-253
Abstract: Using a primer to a conserved nucleotide sequence of previously cloned skin peptides of Phyllomedusa species, two distinct cDNAs were "shotgun" cloned from a skin secretion-derived cDNA library of the frog, Phyllomedusa burmeisteri. The two ORFs separately encode chains A and B of an analog of the previously reported heterodimeric peptide, distinctin. LC-MS/MS analysis of native versus dithiotreitol reduced crude venom, confirmed the predicted primary sequences as well as the cystine link between the two monomers. Distinctin predominantly exists in the venom as a heterodimer (A-B), neither of the constituent peptides were detected as monomer, whereas of the two possible homodimers (A-A or B-B), only B-B was detected in comparatively low quantity. In vitro dimerization of synthetic replicates of the monomers demonstrated that besides heterodimer, both homodimers are also formed in considerable amounts. Distinctin is the first example of an amphibian skin dimeric peptide that is formed by covalent linkage of two chains that are the products of different mRNAs. How this phenomenon occurs in vivo, to exclude significant homodimer formation, is unclear at present but a "favored steric state" type of interaction between chains is most likely. (C) 2012 Elsevier B.V. All rights reserved.
Notes: [Evaristo, Geisa; Pinkse, Martijn; Verhaert, Peter] Delft Univ Technol, Dept Biotechnol, Analyt Biotechnol Grp, NL-2628 BC Delft, Netherlands. [Evaristo, Geisa; Pinkse, Martijn; Verhaert, Peter] Delft Univ Technol, Dept Biotechnol, Netherlands Prote Ctr, NL-2628 BC Delft, Netherlands. [Wang, Lei; Zhou, Mei; Wu, Youjia; Wang, Hui; Chen, Tianbao; Shaw, Chris] Queens Univ Belfast, Sch Pharm, Lab Nat Drug Discovery, Belfast, Antrim, North Ireland. [Verhaert, Peter] Hasselt Univ, Biomed Res Ctr, Diepenbeek, Belgium.
Keywords: Biochemical Research Methods;Amphibian skin peptides; cDNA cloning; Heterodimer; Homodimer; Intramolecular and intermolecular; disulfide bonds; Peptidomics
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ISSN: 1874-3919
e-ISSN: 1876-7737
DOI: 10.1016/j.jprot.2012.09.016
ISI #: 000316706800020
Category: A1
Type: Journal Contribution
Validations: ecoom 2014
Appears in Collections:Research publications

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