Sequence determination by MALDI-TOF mass spectrometry of an insecticidal lentil peptide of the PA1b type

From National Research Council Canada

DOIResolve DOI: http://doi.org/10.1016/j.phytol.2015.03.003
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Name affiliation
  1. National Research Council Canada. Aquatic and Crop Resource Development
TypeArticle
Journal titlePhytochemistry Letters
ISSN1874-3900
Volume12
Pages105–112; # of pages: 8
Subjectarginine; asparagine; aspartic acid; chymotrypsin; cysteine; disulfide; flavone; genomic DNA; glutamic acid; lentil extract; microsomal aminopeptidase; octapeptide; PA1b peptide; peptide; plant extract; proline; pronase; quinestrol; serine; trypsin; tyrosine; alkylation; amino acid sequence; amino terminal sequence; carboxy terminal sequence; collisionally activated dissociation; high performance liquid chromatography; hydrolysis; legume; lentil; matrix assisted laser desorption ionization time of flight mass spectrometry; nucleotide sequence; open reading frame; plant gene; soybean; Glycine max; Lens culinaris; Pisum sativum
Abstract
Mature seeds of lentil (Lens culinaris Medik.) were previously reported to contain an insecticidal cysteine-rich peptide, likely of the albumin-1 subunit b type. The purpose of this work was to determine the amino acid sequence of this insecticidal lentil peptide in an Eston lentil extract by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), after reduction of the disulfide bridges, alkylation of the cysteine residues and hydrolysis by pronase, trypsin, chymotrypsin and endoproteinase Asp-N. Sequences of key fragments were supported by monoisotopic mass measurements and by sequence ions from collision-induced dissociation (CID) experiments with a MALDI-TOF/TOF analyzer (MS/MS analysis). The new 37 amino acid sequence revealed strong similarities to a histidine-containing pea PA1b peptide and to soybean leginsulins but with a unique segment of RSSA in the middle. The lentil PA1b peptide sequence agreed completely with that derived from a L. culinaris genomic DNA sequence. © 2015 Phytochemical Society of Europe.
Publication date
PublisherElsevier
LanguageEnglish
Peer reviewedYes
NPARC number21275720
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