Mitochondria contain thioredoxin (Trx), a regulatory disulfide protein, and an associated flavoenzyme, NADP/Trx reductase, which provide link to NADPH in the organelle. Unlike animal yeast counterparts, function of Trx plant mitochondria is largely unknown. Accordingly, we have applied recently devised proteomic approaches identify soluble Trx-linked proteins isolated from photosynthetic (pea spinach leaves) heterotrophic (potato tubers) sources. Application mitochondrial extracts mutant...

The male gametophyte of Arabidopsis is a three-celled pollen grain that thought to contain almost all the mRNAs needed for germination and rapid tube growth. We generated reference map mature proteome by using multiple protein extraction techniques followed 2-DE ESI-MS/MS. identified 135 distinct proteins from total 179 spots. found half are involved in metabolism (20%), energy generation (17%), or cell structure (12%); these percentages similar those determined transcriptome this similarity...

Wheat flour is one of the world's major food ingredients, in part because unique end-use qualities conferred by abundant glutamine- and proline-rich gluten proteins. Many wheat proteins also present dietary problems for consumers with celiac disease or allergies. Despite importance these it has been particularly challenging to use MS/MS distinguish many a sample relate them gene sequences.

A combined two-dimensional gel electrophoresis-mass spectrometry approach was utilized to identify over 250 proteins of wheat (Triticum aestivum L., cv. Butte 86) starchy endosperm that participate in 13 biochemical processes: ATP interconversion reactions, carbohydrate metabolism, cell division, cytoskeleton, lipid nitrogen protein synthesis/assembly, turnover, signal transduction, storage, stress/defense, transcription/translation, and transport. Endosperm populations were compared at...

A growing number of processes throughout biology are regulated by redox via thiol–disulfide exchange. This mechanism is particularly widespread in plants, where almost 200 proteins have been linked to thioredoxin (Trx), a widely distributed small regulatory disulfide protein. The current study extends regulation Trx amyloplasts, organelles prevalent heterotrophic plant tissues that, among other biosynthetic activities, catalyze the synthesis and storage copious amounts starch. Using...

Abstract Germination of cereals is accompanied by extensive change in the redox state seed proteins. Proteins present oxidized form dry seeds are converted to reduced following imbibition. Thioredoxin (Trx) appears play a role this transition cereals. It not known, however, whether Trx-linked changes restricted or they take place more broadly germinating seeds. To gain information on point, we have investigated model legume, Medicago truncatula. Two complementary gel-based proteomic...

By contrast to chloroplasts, our knowledge of amyloplasts—organelles that synthesize and store starch in heterotrophic plant tissues—is a formative stage. While understanding what is considered their primary function, i.e. the biosynthesis degradation starch, has increased dramatically recent years, relatively little known about other biochemical processes taking place these organelles. To help fill this gap, proteomic analysis amyloplasts isolated from starchy endosperm wheat seeds (10 d...

Abstract Background Flour quality is largely determined by the gluten proteins, a complex mixture of proteins consisting high molecular weight-glutenin subunits (HMW-GS), low (LMW-GS), and α-, γ-, ω-gliadins. Detailed proteomic analyses effects fertilizer temperature on individual gliadin glutenin protein levels are needed to determine how these environmental factors influence flour quality. Results Wheat plants ( Triticum aestivum L. cv. Butte 86) were grown in greenhouses under moderate...

Wheat grains accumulate a variety of low molecular weight proteins that are inhibitors alpha-amylases and proteases play an important protective role in the grain. These have more balanced amino acid compositions than major wheat gluten contribute reserves for both seedling growth human nutrition. The alpha-amylase/protease also interest because they cause IgE-mediated occupational food allergies thereby impact health.The complement genes encoding expressed US bread Butte 86 was...

Application of a thiol‐specific probe, monobromobimane, with proteomics and enzyme assays led to the identification 23 thioredoxin targets in starchy endosperm mature wheat seeds ( Triticum aestivum cv. Butte), almost all containing at least two conserved cysteines. The identified targets, 12 not known be thioredoxin‐linked, function spectrum processes: metabolism (12 targets), protein storage (three), oxidative stress degradation (two), assembly/folding (one) unknown reactions (two). In...

Methods to sequentially extract and fractionate wheat flour proteins were evaluated reliably quantify gliadins, glutenins, albumins/globulins in single samples. Compositions of the resulting protein fractions analyzed by RP-HPLC combined with SDS−PAGE. Unknown identified mass spectrometry or N-terminal sequencing. The best separation recovery discrete albumin/globulin, gliadin, glutenin from same sample was achieved extraction 0.3 M NaI 7.5% 1-propanol followed 2% SDS, 25 mM DTT TRIS, pH...

Certain wheat gluten proteins form large protein polymers that are extractable in 0.5% SDS only after sonication. Although there is a strong relationship between the amounts of these flour and bread-making quality, components have not been thoroughly investigated.Flour from US bread Butte 86 were extracted using two-step procedure with without Proteins further separated by size exclusion chromatography (SEC) into monomeric polymeric fractions analyzed quantitative two-dimensional gel...

While the antigenic specificity and pathogenic relevance of immunologic reactivity to gluten in celiac disease have been extensively researched, immune response nongluten proteins wheat has not characterized. We aimed investigate level molecular antibody disease. Serum samples from patients controls were screened for IgG IgA a protein extract cultivar Triticum aestivum Butte 86. Antibodies further analyzed specific by two-dimensional gel electrophoresis immunoblotting. Immunoreactive...

ABSTRACT ω‐Gliadins were purified from wheat ( Triticum aestivum L. ‘Butte’) flour and characterized. Although reversed‐phase HPLC (RP‐HPLC) separated the 1B‐encoded ω‐gliadins into two fractions, 1B1 1B2, these fractions had nearly identical amino acid compositions, three similar protein bands in SDS‐PAGE, 10 spots two‐dimensional PAGE, N‐terminal sequences. The main components a range mass of 48,900–51,500 when estimated by spectrometry, significantly less than SDS‐PAGE. 1B digested with...

A KCl-soluble, albumin/globulin fraction of wheat (Triticum aestivum L.) starchy endosperm was further separated into a methanol-insoluble that contained metabolic proteins and methanol-soluble “chloroform-methanol” or CM-like proteins. Reduction the disulfide bonds CM with thioredoxin dithiothreitol altered their properties so that, like proteins, they were insoluble in methanol. Glutathione had little effect, indicating dithiol specificity. Proteomic analysis protein revealed presence...

We carried out a parallel transcriptional and proteomic comparison of seeds from transformed bread wheat line that overexpresses transgenic low molecular weight glutenin subunit gene relative to the corresponding nontransformed genotype. Proteomic analyses showed that, during seed development, several classes endosperm proteins were differentially accumulated in endosperm. As result strong increase amount protein, endogenous subunit, all subclasses gliadins, metabolic as well...

Mineral nutrition during wheat grain development has large effects on flour protein content and composition, which in turn affect quality immunogenic potential for a commodity of great economic value. However, it been difficult to define the precise mineral composition because complexity proteome. Recent improvements identification proteins by tandem mass spectrometry (MS/MS) availability comprehensive proteome map from US Butte 86 now make possible document changes proportions individual...