title: Shotgun proteome analysis of honeybee venom using targeted enrichment strategies.
abstract: The aim of this study was to explore the honeybee venom proteome applying a shotgun proteomics approach using different enrichment strategies (combinatorial peptide ligand libraries and solid phase extraction). The studies were conducted using nano-LC/MALDI-TOF/TOF-MS system. The MS analysis of peptide profiles (in the range of 900-4500 Da) and virtual gel-image of proteins from Lab-on-Chip assay (in the range of 10-250 kDa) confirm that use of targeted enrichment strategies increase detection of honeybee venom components. The gel-free shotgun strategy and sophisticated instrumentation led to a significant increase of the sensitivity and higher number of identified peptides in honeybee venom samples, comparing with the current literature. Moreover, 11 of 12 known honeybee venom allergens were acknowledged and 4 new, so far uncharacterized proteins were identified. In addition, similarity searches were performed in order to investigate biological relations and homology between newly identified proteins sequences from Apis mellifera and other Hymenoptera.
authors.affiliation: Department of Inorganic & Analytical Chemistry, Poznan University of Medical Sciences, 6 Grunwaldzka Street, 60-780 Poznań, Poland. Electronic address: jmatysiak@ump.edu.pl.
authors.forename: Jan
authors.initials: J
authors.lastname: Matysiak
authors.affiliation: Department of Inorganic & Analytical Chemistry, Poznan University of Medical Sciences, 6 Grunwaldzka Street, 60-780 Poznań, Poland.
authors.forename: Joanna
authors.initials: J
authors.lastname: Hajduk
authors.affiliation: Department of Inorganic & Analytical Chemistry, Poznan University of Medical Sciences, 6 Grunwaldzka Street, 60-780 Poznań, Poland.
authors.forename: Łukasz
authors.initials: Ł
authors.lastname: Pietrzak
authors.affiliation: Institute of Pharmacy, Faculty of Natural Sciences I, Martin Luther University Halle-Wittenberg, Halle (Saale), Germany.
authors.forename: Christian E H
authors.initials: CE
authors.lastname: Schmelzer
authors.affiliation: Department of Inorganic & Analytical Chemistry, Poznan University of Medical Sciences, 6 Grunwaldzka Street, 60-780 Poznań, Poland.
authors.forename: Zenon J
authors.initials: ZJ
authors.lastname: Kokot
mesh_terms: D000818:Animals; D001514:Bee Venoms / Q000737:chemistry*; D001516:Bees; D002853:Chromatography, Liquid; D056656:Lab-On-A-Chip Devices; D020543:Proteome; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D001514:Bee Venoms; D020543:Proteome
keywords: Apis mellifera; Lab-on-Chip; MALDI; Mass spectrometry; Proteomics