The ?Dark Side? of Food Stuff Proteomics: The CPLL-Marshals Investigate

The present review deals with analysis of the proteome of animal and plant-derived food stuff, as well as of non-alcoholic and alcoholic beverages. The survey is limited to those system investigated with the help of combinatorial peptide ligand libraries, a most powerful technique allowing access to low- to very-low-abundance proteins, i.e. to those proteins that might characterize univocally a given biological system and, in the case of commercial food preparations, attest their genuineness or adulteration. Among animal foods the analysis of cow’s and donkey’s milk is reported, together with the proteomic composition of egg white and yolk, as well as of honey, considered as a hybrid between floral and animal origin. In terms of plant and fruits, a survey is offered of spinach, artichoke, banana, avocado, mango and lemon proteomics, considered as recalcitrant tissues in that small amounts of proteins are dispersed into a large body of plant polymers and metabolites. As examples of non-alcoholic beverages, ginger ale, coconut milk, a kola drink, almond milk and orgeat syrup are analyzed. Finally, the trace proteome of white and red wines, beer and aperitifs is reported, with the aim of tracing the industrial manipulations and herbal usage prior to their commercialization. The review ends with a comparison between mammalian and plant proteomics highlighting the difficulties besieging analysis of any vegetable proteome

Protein Equalizer™ Technology*: The quest for a “democratic proteome”

A review. No proteome can be considered “democratic”, but rather “oligarchic”, since a few proteins dominate the landscape and often obliterate the signal of the rare ones. This is the reason why most scientists lament that, in proteome analysis, the same set of abundant proteins is seen again and again. A host of pre-fractionation techniques have been described, but all of them, one way or another, are besieged by problems, in that they are based on a “depletion principle”, i.e. getting rid of the unwanted species. Yet “democracy” calls not for killing the enemy, but for giving “equal rights” to all people. One way to achieve that would be the use of “Protein Equalizer Technology” for reducing protein concentration differences. This comprises a diverse library of combinatorial ligands coupled to spherical porous beads. When these beads come into contact with complex proteomes (e.g. human urine and serum, egg white, and any cell lysate, for that matter) of widely differing protein composition and relative abundances, they are able to “equalize” the protein population, by sharply reducing the concentration of the most abundant components, while simultaneously enhancing the concentration of the most dilute species. It is felt that this novel method could offer a strong step forward in bringing the “unseen proteome” (due to either low abundance and/or presence of interference) within the detection capabilities of current proteomics detection methods. Examples are given of equalization of human urine and serum samples, resulting in the discovery of a host of proteins never reported before. Additionally, these beads can be used to remove host cell proteins from purified recombinant proteins or protein purified from natural sources that are intended for human consumption. These proteins typically reach purities of the order of 98%: higher purities often become prohibitively expensive. Yet, if incubated with “equalizer beads”, these last impurities can be effectively removed at a small cost and with minute losses of the main, valuable product

“Proteomics approaches for studying chemoresistance in cancer”

The role of various proteins involved in drug resistance in tumor cells is discussed in this review. Two types of studies are covered: those performed in the preproteomics era and those carried out with modern proteomic tools, namely 2D (electrophoretic) maps and 2D chromatography. In the preproteomic studies, one protein had generally been held responsible for a given chemoresistance. However, analysis via proteomic tools may reveal entire sets of proteins that are up- or downregulated (or switched on/off) in chemoresistant tumor cell lines compared with parental tumor lines. Therefore, it appears more realistic to expect that exposure of cells to drugs results in the activation of different mechanisms of resistance. Such investigations have led to the broadly shared opinion that exposure of cells to drugs results in the activation of different mechanisms of resistance, and that a specific drug-resistant phenotype consists of several molecular mechanisms that are simultaneously active. The proteomic papers reviewed clearly support the hypothesis that many metabolic pathways are affected during the resistance process. Although the modulation of expression levels of such proteins is not clear proof of their role in drug resistance per se, at least some of the themes are very likely to be involved in the resistance phenotype, and thus may be potential targets for new drugs. It is hoped that this review will bring new insight in this field and will stimulate novel and deeper searches with proteomic tools (including prefractionation of subcellular organelles, such as nuclei, to bring to the fore low-abundance proteins that might be responsible for the onset of drug resistance)

In taberna quando sumus: a drunkard's cakewalk through wine proteomics

A review. Anal. of white and red wine trace proteomes via capture with combinatorial peptide ligand libraries (CPLL) is reported here. Most of the alc. beverages tested (all of Italian origin) were found to contain only traces of casein (on av. from 20 to 60 μg/L, with a detectability of as low as 1 μg/L) and not any grape protein any longer, as they had been fined with bovine casein (surprisingly also red wines for which the typical fining agent is egg albumin). However, anal. of untreated white wine (Recite, from Garganega grapes in the Veneto region) via CPLL capture indeed permitted to detect close to 100 unique gene products from the grapes, suggesting the possibility of proteotyping grand crus, i.e. those aged, high quality wines that should not be treated with fining agents. Thus the CPLL technique could become a formidable tool for traceability of beverages in particular and of foodstuff in general. For trace protein anal., a new, most powerful CPLL methodol. emerges: capture at pH=2.2 in 0.1 % trifluoroacetic acid (TFA) under the conditions mimicking reversed-phase mechanisms of adsorption