pr300617a_si_001.pdf (1.26 MB)
Download fileSize-Selective Fractionation and Visual Mapping of Allergen Protein Chemistry in Arachis hypogaea
journal contribution
posted on 2012-11-02, 00:00 authored by Christine M. Hebling, Mark M. Ross, John H. Callahan, Melinda
A. McFarlandPeanuts (Arachis hypogaea) in addition
to milk,
eggs, fish, crustaceans, wheat, tree nuts, and soybean are commonly
referred to as the “big eight” foods that contribute
to the majority of food allergies worldwide. Despite the severity
of allergic reactions and growing prevalence in children and adults,
there is no cure for peanut allergy, leaving avoidance as the primary
mode of treatment. To improve analytical methods for peanut allergen
detection, researchers must overcome obstacles involved in handling
complex food matrices while attempting to decipher the chemistry that
underlies allergen protein interactions. To address such challenges,
we conducted a global proteome characterization of raw peanuts using
a sophisticated GELFrEE-PAGE-LC–MS/MS platform consisting of
gel-based protein fractionation followed by mass spectrometric identification.
The in-solution mass-selective protein fractionation: (1) enhances
the number of unique peptide identifications, (2) provides a visual
map of protein isoforms, and (3) aids in the identification of disulfide-linked
protein complexes. GELFrEE-PAGE-LC–MS/MS not only overcomes
many of the challenges involved in the study of plant proteomics,
but enriches the understanding of peanut protein chemistry, which
is typically unattainable in a traditional bottom-up proteomic analysis.
A global understanding of protein chemistry in Arachis hypogaea ultimately will aid the development of improved methods for allergen
detection in food.