title: Computer aided identification of a Hevein-like antimicrobial peptide of bell pepper leaves for biotechnological use.
abstract: Antimicrobial peptides from plants present mechanisms of action that are different from those of conventional defense agents. They are under-explored but have a potential as commercial antimicrobials. Bell pepper leaves ('Magali R') are discarded after harvesting the fruit and are sources of bioactive peptides. This work reports the isolation by peptidomics tools, and the identification and partially characterization by computational tools of an antimicrobial peptide from bell pepper leaves, and evidences the usefulness of records and the in silico analysis for the study of plant peptides aiming biotechnological uses.
  
  
  
  Aqueous extracts from leaves were enriched in peptide by salt fractionation and ultrafiltration. An antimicrobial peptide was isolated by tandem chromatographic procedures. Mass spectrometry, automated peptide sequencing and bioinformatics tools were used alternately for identification and partial characterization of the Hevein-like peptide, named HEV-CANN. The computational tools that assisted to the identification of the peptide included BlastP, PSI-Blast, ClustalOmega, PeptideCutter, and ProtParam; conventional protein databases (DB) as Mascot, Protein-DB, GenBank-DB, RefSeq, Swiss-Prot, and UniProtKB; specific for peptides DB as Amper, APD2, CAMP, LAMPs, and PhytAMP; other tools included in ExPASy for Proteomics; The Bioactive Peptide Databases, and The Pepper Genome Database. The HEV-CANN sequence presented 40 amino acid residues, 4258.8 Da, theoretical pI-value of 8.78, and four disulfide bonds. It was stable, and it has inhibited the growth of phytopathogenic bacteria and a fungus. HEV-CANN presented a chitin-binding domain in their sequence. There was a high identity and a positive alignment of HEV-CANN sequence in various databases, but there was not a complete identity, suggesting that HEV-CANN may be produced by ribosomal synthesis, which is in accordance with its constitutive nature.
  
  
  
  Computational tools for proteomics and databases are not adjusted for short sequences, which hampered HEV-CANN identification. The adjustment of statistical tests in large databases for proteins is an alternative to promote the significant identification of peptides. The development of specific DB for plant antimicrobial peptides, with information about peptide sequences, functional genomic data, structural motifs and domains of molecules, functional domains, and peptide-biomolecule interactions are valuable and necessary.
authors.affiliation: Department of Biochemistry and Molecular Biology, Universidade Federal de Viçosa, Viçosa, MG, 36570-900, Brazil.
authors.forename: Patrícia Dias
authors.initials: PD
authors.lastname: Games
authors.affiliation: Department of Biochemistry and Molecular Biology, Universidade Federal de Viçosa, Viçosa, MG, 36570-900, Brazil.
authors.forename: Elói Quintas Gonçalves
authors.initials: EQ
authors.lastname: daSilva
authors.affiliation: Department of Biochemistry and Molecular Biology, Universidade Federal de Viçosa, Viçosa, MG, 36570-900, Brazil.
authors.forename: Meire de Oliveira
authors.initials: MO
authors.lastname: Barbosa
authors.affiliation: Department of Biochemistry and Molecular Biology, Universidade Federal de Viçosa, Viçosa, MG, 36570-900, Brazil.
authors.forename: Hebréia Oliveira
authors.initials: HO
authors.lastname: Almeida-Souza
authors.affiliation: Department of Biochemistry and Molecular Biology, Universidade Federal de Viçosa, Viçosa, MG, 36570-900, Brazil.
authors.forename: Patrícia Pereira
authors.initials: PP
authors.lastname: Fontes
authors.affiliation: Department of Biochemistry and Molecular Biology, Universidade Federal de Viçosa, Viçosa, MG, 36570-900, Brazil.
authors.forename: Marcos Jorge
authors.initials: MJ
authors.lastname: deMagalhães
authors.affiliation: Department of Plant Science, Universidade Federal de Viçosa, Viçosa, MG, 36570-900, Brazil.
authors.forename: Paulo Roberto Gomes
authors.initials: PR
authors.lastname: Pereira
authors.affiliation: Embrapa Genetic Resources & Biotechnology, Brazilian Agricultural Research Corporation, Brasília, DF, 70770-900, Brazil.
authors.forename: Maura Vianna
authors.initials: MV
authors.lastname: Prates
authors.affiliation: Department of Biochemistry and Immunology-ICB, Universidade Federal de Minas Gerais, Av. Antônio Carlos 6627, Belo Horizonte, MG, 31270-901, Brazil.
authors.forename: Gloria Regina
authors.initials: GR
authors.lastname: Franco
authors.affiliation: Department of Computer Science-ICEX, Universidade Federal de Minas Gerais, Av. Antônio Carlos 6627, Belo Horizonte, MG, 31270-901, Brazil.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Faria-Campos
authors.affiliation: Department of Computer Science-ICEX, Universidade Federal de Minas Gerais, Av. Antônio Carlos 6627, Belo Horizonte, MG, 31270-901, Brazil.
authors.forename: Sérgio Vale Aguiar
authors.initials: SV
authors.lastname: Campos
authors.affiliation: Department of Biochemistry and Molecular Biology, Universidade Federal de Viçosa, Viçosa, MG, 36570-900, Brazil. baracat@ufv.br.
authors.forename: Maria Cristina
authors.initials: MC
authors.lastname: Baracat-Pereira
mesh_terms: D000595:Amino Acid Sequence; D000890:Anti-Infective Agents / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology; D023181:Antimicrobial Cationic Peptides / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology; D001709:Biotechnology; D002212:Capsicum / Q000737:chemistry* / Q000378:metabolism; D056148:Chromatography, Reverse-Phase; D030562:Databases, Protein; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D010455:Peptides / Q000032:analysis / Q000302:isolation & purification; D018515:Plant Leaves / Q000737:chemistry* / Q000378:metabolism; D037121:Plant Lectins / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology; D010940:Plant Proteins / Q000032:analysis / Q000378:metabolism; D040901:Proteomics; D016415:Sequence Alignment
chemical_list: D000890:Anti-Infective Agents; D023181:Antimicrobial Cationic Peptides; D010455:Peptides; D037121:Plant Lectins; D010940:Plant Proteins; C045875:hevein
keywords: Antimicrobial peptide; Bell pepper; Bioinformatics; Biotechnology; Computational tools; Hevein-like; Peptidomics; Plant defense