title: Conformational and functional effects induced by D- and L-amino acid epimerization on a single gene encoded peptide from the skin secretion of Hypsiboas punctatus.
abstract: Skin secretion of Hypsiboas punctatus is the source of a complex mixture of bioactive compounds where peptides and small proteins prevail, similarly to many other amphibians. Among dozens of molecules isolated from H. punctatus in a proteomic based approach, we report here the structural and functional studies of a novel peptide named Phenylseptin (FFFDTLKNLAGKVIGALT-NH2) that was purified as two naturally occurring D- and L-Phes configurations. The amino acid epimerization and C-terminal amidation for both molecules were confirmed by a combination of techniques including reverse-phase UFLC, ion mobility mass spectrometry, high resolution MS/MS experiments, Edman degradation, cDNA sequencing and solid-phase peptide synthesis. RMSD analysis of the twenty lowest-energy (1)H NMR structures of each peptide revealed a major 90° difference between the two backbones at the first four N-terminal residues and substantial orientation changes of their respective side chains. These structural divergences were considered to be the primary cause of the in vitro quantitative differences in antimicrobial activities between the two molecules. Finally, both molecules elicited equally aversive reactions in mice when delivered orally, an effect that depended entirely on peripheral gustatory pathways.
authors.affiliation: Laboratório de Espectrometria de Massa, Embrapa Recursos Genéticos e Biotecnologia, Brasília-Distrito Federal, Brasil.
authors.forename: Mariana T Q
authors.initials: MT
authors.lastname: de Magalhães
authors.forename: Eder A
authors.initials: EA
authors.lastname: Barbosa
authors.forename: Maura V
authors.initials: MV
authors.lastname: Prates
authors.forename: Rodrigo M
authors.initials: RM
authors.lastname: Verly
authors.forename: Victor Hugo O
authors.initials: VH
authors.lastname: Munhoz
authors.forename: Ivan E
authors.initials: IE
authors.lastname: de Araújo
authors.forename: Carlos
authors.initials: C
authors.lastname: Bloch
mesh_terms: D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000737:chemistry*; D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D001001:Anura / Q000235:genetics*; D001483:Base Sequence; D008297:Male; D013058:Mass Spectrometry; D051379:Mice; D018345:Mice, Knockout; D008826:Microbial Sensitivity Tests; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D010455:Peptides / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D011487:Protein Conformation; D016415:Sequence Alignment; D012867:Skin / Q000378:metabolism*; D050053:TRPM Cation Channels / Q000172:deficiency; D013649:Taste; D055697:Taste Perception / Q000235:genetics
chemical_list: D000596:Amino Acids; D023181:Antimicrobial Cationic Peptides; D010455:Peptides; D050053:TRPM Cation Channels; C498617:Trpm5 protein, mouse
references.citation: J Exp Med. 1968 Nov 1;128(5):877-94
references.pmid: 4176225
references.citation: CNS Spectr. 2011 Feb;16(2):63
references.pmid: 22789775
references.citation: Peptides. 2005 Apr;26(4):565-73
references.pmid: 15752569
references.citation: FASEB J. 2007 Aug;21(10):2466-73
references.pmid: 17384140
references.citation: BMJ. 2000 Dec 23-30;321(7276):1591-4
references.pmid: 11124191
references.citation: J Virol Methods. 2006 Nov;137(2):280-6
references.pmid: 16890998
references.citation: Ciba Found Symp. 1994;186:77-85; discussion 85-90
references.pmid: 7768159
references.citation: Biol Lett. 2005 Dec 22;1(4):500-3
references.pmid: 17148243
references.citation: Lancet. 1997 May 31;349(9065):1569-81
references.pmid: 9174558
references.citation: J Am Soc Mass Spectrom. 2005 Feb;16(2):158-65
references.pmid: 15694766
references.citation: Curr Protoc Bioinformatics. 2002 Aug;Chapter 2:Unit 2.3
references.pmid: 18792934
references.citation: Biochim Biophys Acta. 1999 Dec 15;1462(1-2):55-70
references.pmid: 10590302
references.citation: J Chem Ecol. 2004 Nov;30(11):2215-30
references.pmid: 15672666
references.citation: Anim Behav. 1999 Nov;58(5):921-931
references.pmid: 10564594
references.citation: Proc Natl Acad Sci U S A. 2010 Dec 7;107(49):21205-10
references.pmid: 21088219
references.citation: Nucleic Acids Res. 2005 Jan 1;33(Database issue):D192-6
references.pmid: 15608175
references.citation: J Biol Chem. 1994 Dec 16;269(50):31635-41
references.pmid: 7989335
references.citation: J Biomol NMR. 2009 Aug;44(4):213-23
references.pmid: 19548092
references.citation: Proc Natl Acad Sci U S A. 2011 Jul 5;108(27):11235-40
references.pmid: 21690383
references.citation: J Mol Biol. 2006 Feb 3;355(5):879-86
references.pmid: 16343537
references.citation: Peptides. 1994;15(2):199-202
references.pmid: 8008623
references.citation: J Biol Chem. 2004 Mar 26;279(13):13018-26
references.pmid: 14715660
references.citation: J Biol Chem. 2002 Dec 20;277(51):49332-40
references.pmid: 12379643
references.citation: Proc Natl Acad Sci U S A. 1996 Apr 30;93(9):4036-9
references.pmid: 8633012
references.citation: Peptides. 2004 Jun;25(6):1035-54
references.pmid: 15203252
references.citation: Biopolymers. 1998;47(6):435-50
references.pmid: 10333736
references.citation: Biochemistry. 2006 Apr 4;45(13):4266-76
references.pmid: 16566601
references.citation: Cell. 2003 Feb 7;112(3):293-301
references.pmid: 12581520
references.citation: Peptides. 2006 Sep;27(9):2137-46
references.pmid: 16797783
references.citation: Proc Natl Acad Sci U S A. 1990 Jul;87(14):5509-13
references.pmid: 2196570
references.citation: J Magn Reson. 2001 Apr;149(2):239-44
references.pmid: 11318623
references.citation: Kaku Igaku. 1981 Jun;18(5):601-8
references.pmid: 6796734
references.citation: Biochem Biophys Res Commun. 1992 Nov 30;189(1):184-90
references.pmid: 1449472
references.citation: J Agric Food Chem. 2006 Dec 27;54(26):10102-11
references.pmid: 17177547
references.citation: J Mol Biol. 1993 Jun 20;231(4):1049-67
references.pmid: 8515464
references.citation: Protein Sci. 1992 Jun;1(6):736-51
references.pmid: 1304915
references.citation: Biochem Biophys Res Commun. 2006 Sep 1;347(3):739-46
references.pmid: 16844081
references.citation: Proc Natl Acad Sci U S A. 1989 Jul;86(13):5188-92
references.pmid: 2544892
references.citation: Eur J Cell Biol. 2006 Jan;85(1):25-34
references.pmid: 16373172
references.citation: Neurotherapeutics. 2009 Jan;6(1):43-52
references.pmid: 19110198
references.citation: Biochem Biophys Res Commun. 1989 May 15;160(3):1015-20
references.pmid: 2597281
references.citation: Cell Mol Neurobiol. 1995 Aug;15(4):465-93
references.pmid: 8565049
references.citation: Proc Natl Acad Sci U S A. 1987 Aug;84(15):5449-53
references.pmid: 3299384
references.citation: J Am Vet Med Assoc. 1993 Jan 15;202(2):229-49
references.pmid: 8428829
references.citation: Nature. 2006 Mar 9;440(7081):208-11
references.pmid: 16525472
references.citation: Nature. 1951 Jun 23;167(4260):1033
references.pmid: 14843163
references.citation: Toxicon. 1993 Sep;31(9):1099-111
references.pmid: 8266343
references.citation: Int J Pept Protein Res. 1992 Mar;39(3):258-64
references.pmid: 1399265
references.citation: Eur J Pharmacol. 1989 Mar 14;162(1):123-8
references.pmid: 2542051
references.citation: EXS. 1998;85:57-71
references.pmid: 9949868