Thioredoxins are 12-kDa proteins functional in the regulation of cellular processes throughout the animal, plant, and microbial kingdoms. Growing evidence with seeds suggests that an h -type of thioredoxin, reduced by NADPH via NADP-thioredoxin reductase, reduces disulfide bonds of target proteins and thereby acts as a wakeup call in germination. A better understanding of the role of thioredoxin in seeds as well as other systems could be achieved if more were known about the target proteins. To this end, we have devised a strategy for the comprehensive identification of proteins targeted by thioredoxin. Tissue extracts incubated with reduced thioredoxin are treated with a fluorescent probe (monobromobimane) to label sulfhydryl groups. The newly labeled proteins are isolated by conventional two-dimensional electrophoresis: ( i ) nonreducing/reducing or ( ii ) isoelectric focusing/reducing SDS/PAGE. The isolated proteins are identified by amino acid sequencing. Each electrophoresis system offers an advantage: the first method reveals the specificity of thioredoxin in the reduction of intramolecular vs. intermolecular disulfide bonds, whereas the second method improves the separation of the labeled proteins. By application of both methods to peanut seed extracts, we isolated at least 20 thioredoxin targets and identified 5—three allergens (Ara h2, Ara h3, and Ara h6) and two proteins not known to occur in peanut (desiccation-related and seed maturation protein). These findings open the door to the identification of proteins targeted by thioredoxin in a wide range of systems, thereby enhancing our understanding of its function and extending its technological and medical applications.

In the era of SDGs, useful plants which provide valuable industrial outputs and at the same time pose less impact on the environment should be explored. Hemp seems one of the most relevant gluten-free crop plants to meet such requirements. Its high nutritional value is comparable to soy. Moreover, almost the whole body of the hemp plant has a wide array of utility: industrial production of food, fiber, and construction materials. In view of environmental sustainability, hemp requires less pesticides or water in cultivation compared to cotton, a representative fiber plant. This short review investigates hemp’s sustainability as a plant as well as its utility value as a highly nutritional material in the food industry. Recent application research of hemp protein in food processing includes plant milk, emulsifiers, fortification of gluten-free bread, plant-based meat production, as well as membrane formation. These studies have revealed distinctive properties of hemp protein, especially in relation to disulfide (S-S)/sulfhydryl (-SH)-mediated interactions with protein from other sources. While its cultivation area and industrial use were limited for a while over confusion with marijuana, the market for industrial hemp is growing rapidly because it has been highly reevaluated in multiple areas of industry. Conclusively, with its sustainability as a plant as well as its distinctive useful property of the seed protein, hemp has promising value in the development of new foods.

The mobilization of storage proteins (glutelins) in germinating rice seeds was accompanied by an ordered sequential combination of proteolysis and reduction of disulfide groups. Mobilization was followed by application of non-reducing/reducing two dimensional-PAGE after monobromobimane labeling of the sulfhydryl groups of the proteins in intact seeds.