Osmotic stress is accompanied by protein glycation inArabidopsis thaliana

Abstract

HighlightOsmotic stress enhances the rate of protein glycation and monosaccharide autoxidation is the main pathway.

Mining seed proteome: from protein dynamics to modification profiles

“…Among many others, Progenta anionic acid labile surfactant (AALSII) is another option for enzymatic digestion of hydrophobic (e.g., membrane) proteins (Waas et al, 2016). We recently used this detergent for analysis of the total Arabidopsis leaf proteome Paudel et al, 2016). Later, we successfully applied this approach to germinating seeds of oilseed rape (Brassica napus) .…”

“…After several cleaning steps, proteins are precipitated from phenol phase with ice-cold methanolic ammonium acetate solution, sequentially washed with methanol and acetone, dried and completely reconstituted in a "shotgun buffer" containing haotropic agents (urea and thiourea) and detergent (Progenta Anionic Acid-Labile Surfactant, AALSII) . Afterwards, after determination of protein contents and verification of results with SDS-PAGE, solubilized proteins are digested by trypsin (Greifenhagen et al, 2016). When completeness of the proteolysis is verified with SDS-PAGE, AALS can be destroyed and the digests can be pre-cleaned with RP-SPE either in a cartridge or stage-tip format as described by .…”

“…Indeed, as only a limited number of quasi-molecular ions can be involved in fragmentation in each DDA cycle, an essential (if not the major) part of a proteome is left undetected. To avoid (or at least minimize) this problem, LC-MS analysis needs to rely either on enrichment/pre-fractionation techniques (when annotation of the maximum possible number of proteins is the main scope of the study) or prolonged gradients (when reliable label-free quantification is desired) (Bollineni, Hoffmann, and Fedorova, 2011;Paudel et al, 2016 ). The choice of mass analyzer is based on the principle goal of the research.…”

“…As was mentioned before, the coverage of non-enzymatic modifications (which are typically represented with low-intensity signals in chromatograms) can be increased by implementation of enrichment/depletion and pre-fractionation procedures (Bilova et al, 2016b;Paudel et al, 2016). Besides, fragmentation of highintensity signals of abundant unmodified peptides can be suppressed in DDA experiments by using the resultbased exclusion strategy .…”

Quantification of prospective type 2 diabetes mellitus biomarkers by stable isotope dilution with bi-labeled standard glycated peptides

“…15 Moreover, the existence of structural consensus moieties cannot be excluded, as glycation might affect the protein structure. 39 In our study we applied QqTOF-MS as a detection method. Although triple quadrupole (QqQ) -MS employing multiple reaction monitoring (MRM) is considered to be a gold standard in quantication of glycation products, 19 the QqTOF approach (as the variant of HR-MS) seems to be more applicable for the step of the multi-biomarker method development.…”