MS^E Based Multiplex Protein Analysis Quantified Important Allergenic Proteins and Detected Relevant Peptides Carrying Known Epitopes in Wheat Grain Extracts

The amount of clinically relevant, allergy-related proteins in wheat grain is still largely unknown. The application of proteomics may create a platform not only for identification and characterization, but also for quantitation of these proteins. The aim of this study was to evaluate the data-independent quantitative mass spectrometry (MS^E) approach in combination with 76 wheat allergenic sequences downloaded from the AllergenOnline database (www.allergenonline.org) as a starting point. Alcohol soluble extracts of gliadin and glutenin proteins were analyzed. This approach has resulted in identification and quantification of 15 allergenic protein isoforms that belong to amylase/trypsin inhibitors, γ-gliadins, and high or low molecular weight glutenins. Additionally, several peptides carrying four previously discovered epitopes of γ-gliadin B precursor have been detected. These data were validated against the UniProt database, which contained 11764 <i>Triticeae</i> protein sequences. The identified allergens are discussed in relation to Baker’s asthma, food allergy, wheat dependent exercise induced anaphylaxis, atopic dermatitis, and celiac disease (i.e., gluten-sensitive enteropathy). In summary, the results showed that the MS^E approach is suitable for quantitative analysis and allergens profiling in wheat varieties and/or other food matrices

Soybeans Grown in the Chernobyl Area Produce Fertile Seeds that Have Increased Heavy Metal Resistance and Modified Carbon Metabolism

<div><p>Plants grow and reproduce in the radioactive Chernobyl area, however there has been no comprehensive characterization of these activities. Herein we report that life in this radioactive environment has led to alteration of the developing soybean seed proteome in a specific way that resulted in the production of fertile seeds with low levels of oil and β-conglycinin seed storage proteins. Soybean seeds were harvested at four, five, and six weeks after flowering, and at maturity from plants grown in either non-radioactive or radioactive plots in the Chernobyl area. The abundance of 211 proteins was determined. The results confirmed previous data indicating that alterations in the proteome include adaptation to heavy metal stress and mobilization of seed storage proteins. The results also suggest that there have been adjustments to carbon metabolism in the cytoplasm and plastids, increased activity of the tricarboxylic acid cycle, and decreased condensation of malonyl-acyl carrier protein during fatty acid biosynthesis.</p> </div

Establishing a Leaf Proteome Reference Map for <i>Ginkgo biloba</i> Provides Insight into Potential Ethnobotanical Uses

Although ginkgo (Maidenhair tree, Ginkgo biloba L.) is an ancient medicinal and ornamental tree, there has not previously been any systematic proteomic study of the leaves. Herein we describe results from the initial study identifying abundant ginkgo leaf proteins and present a gel reference map. Proteins were isolated from fully developed mature leaves in biological triplicate and analyzed by two-dimensional electrophoresis plus tandem mass spectrometry. Using this approach, we were able to reproducibly quantify 190 abundant protein spots, from which 157 proteins were identified. Most of identified proteins are associated with the energy and protein destination/storage categories. The reference map provides a basis for understanding the accumulation of flavonoids and other phenolic compounds in mature leaves (e.g., identification of chalcone synthase, the first committed enzyme in flavonoid biosynthesis). We additionally detected several proteins of as yet unknown function. These proteins comprise a pool of potential targets that might be useful in nontraditional medical applications

Functional classification of the 211 soybean proteins with paired abundances between soybean seed development in non-radioactive and radioactive Chernobyl fields.

<p>The most abundant functional class was proteins associated with Protein destination and storage followed by Metabolic and Energy proteins.</p

Schematic view of metabolic pathways for carbon assimilation, nitrogen reassimilation, and respiration during soybean seed development in non-radioactive (black) and radioactive (red) Chernobyl areas.

<p>Graphs shows abundance of protein spots as relative volumes. Proteins are displayed on corresponding metabolic pathways. Dashed arrows are used when no protein was detected or was detected only during seed development in one area. Abbreviations for metabolites: ACP, acyl carrier protein; ADP, adenosine diphosphate; ATP, adenosine triphosphate; DHAP, dihydroxyacetone phosphate; F-1,6bisP, fructose 1,6 bis phosphate; F-6-P, fructose 6 phosphate; G-1-P, glucose 1 phosphate; G3P, glyceraldehyde 3-phosphate; PEP, phosphoenolpyruvate; 1,3-bis PGA, 1,3 bis phosphoglyceric acid; 2-PGA, 2 phosphoglyceric acid; 3-PGA, 3 phosphoglyceric acid; R-1,5-bis P, ribulose 1,5 bis phosphate; R-5-P, ribulose 5-phosphate; UDP, uridine diphosphate; UTP, uridine triphosphate; UDP-G, UDP-glucose. Abbreviations for enzymes: E₃ (L), dihydrolipoamide dehydrogenase, L-protein of glycine decarboxylase system; FBA, fructose bisphosphate aldolase; GDH1, glutamate dehydrogenase 1; GS1, glutamine synthetase; KAS1, beta-ketoacyl-ACP synthetase I; LHCB, chlorophyll a/b-binding protein (light-harvesting complex II); MDH, malate dehydrogenase; P, P-protein of glycine decarboxylase system; PEPC, phosphoenolpyruvate carboxylase; PGK, phosphoglycerate kinase; SBP, sucrose binding protein; SuSy, sucrose synthase; SQR, succinate dehydrogenase (ubiquinone); T, T-protein of glycine decarboxylase system; TPI, triose-phosphate isomerase; UDPGP, UTP–glucose-1-phosphate uridylyltransferase.</p

Physiological characterization of developing and dry seeds harvested from non-radioactive and radioactive Chernobyl areas.

<p>A. Seeds were characterized for fresh/dry weight, width, length, and thickness. B. Transfer coefficients (TC) calculated for mature seeds and soybean shoot systems (plants without roots and seed pods) harvested from the radioactive area. C. Total oil content in dry seeds. The standard deviations are shown as error dashes.</p

Composite protein abundance profiles for metabolic classes that were established by summing of abundance profiles for individual proteins within metabolic groups Metabolism and Energy and for storage proteins (β-conglycinins and glycinins) during soybean seed development in radioactive (dashed line) and control Chernobyl areas.

<p>The number of proteins within each composite protein abundance profile is shown.</p