We have reported a set of electrokinetically pumped sheath flow nanoelectrospray interfaces to couple capillary zone electrophoresis with mass spectrometry. A separation is threaded through cross into glass emitter. side arm provides fluidic contact buffer reservoir that connected power supply. The potential applied the drives electro-osmosis in emitter pump fluid at nanoliter per minute rates. Our first-generation interface placed flat-tipped Sensitivity was inversely related orifice size...

14-3-3 proteins function as major regulators of primary metabolism and cellular signal transduction in plants. However, their involvement plant defense stress responses is largely unknown. In order to better address functions the rice 14-3-3/GF14 abiotic responses, we examined GF14 family that comprises eight numbers. The phylogenetic comparison with Arabidopsis revealed majority GF14s might have evolved an independent branch. At least four genes, GF14b, GF14c, GF14e Gf14f were...

Femtogram proteomics: An ultrasensitive capillary zone electrophoresis–mass spectrometry system that is based on an improved nanospray interface has been developed. This used for the analysis of picogram to femtogram amounts E. coli digests; example, over 100 proteins were identified from 16 pg digests by tandem mass spectrometry. AMTs=accurate and time tags. As a service our authors readers, this journal provides supporting information supplied authors. Such materials are peer reviewed may...

We demonstrate the use of capillary zone electrophoresis with an electrokinetically pumped sheath-flow electrospray interface for analysis a tryptic digest sample intermediate protein complexity, secreted fraction Mycobacterium marinum. For electrophoretic analysis, 11 fractions were generated from using reverse-phase liquid chromatography; each was analyzed by CZE-ESI-MS/MS, and 334 peptides corresponding to 140 proteins identified in 165 min mass spectrometer time at 95% confidence (FDR <...

ZnO NPs increased metal nutrient accumulation and reprogrammed carbohydrate amino acid metabolism in tomato plants. They also improved Fe deficiency tolerance by improving accumulation, antioxidative capacity contents of sugars acids.

While there is a rich literature on transcription dynamics during the development of many organisms, protein data limited. We used iTRAQ isotopic labeling and mass spectrometry to generate largest developmental proteomic dataset for any animal. Expression nearly 4,000 proteins Xenopuslaevis was generated from fertilized egg neurula embryo. clusters into groups. The cluster profiles accurately reflect major events that mark changes in gene expression patterns early Xenopus development....

Capillary zone electrophoresis (CZE)-tandem mass spectrometry (MS/MS) has been recognized as a useful tool for top-down proteomics. However, its performance deep proteomics is still dramatically lower than widely used reversed-phase liquid chromatography (RPLC)-MS/MS. We present an orthogonal multidimensional separation platform that couples size exclusion (SEC) and RPLC based protein prefractionation to CZE-MS/MS of Escherichia coli. The generated high peak capacity (∼4000) intact proteins,...

Protein complexes are the functional machines in cell and heterogeneous due to protein sequence variations post-translational modifications (PTMs). Here, we present an automated nondenaturing capillary isoelectric focusing-mass spectrometry (ncIEF-MS) methodology for uncovering microheterogeneity of intact complexes. The method exhibited superior separation resolution than conventional native zone electrophoresis (nCZE-MS). In our study, ncIEF-MS achieved liquid-phase separations MS...

Understanding cancer metastasis at the proteoform level is crucial for discovering previously unknown protein biomarkers diagnosis and drug development. We present first top-down proteomics (TDP) study of a pair isogenic human nonmetastatic metastatic colorectal (CRC) cell lines (SW480 SW620). identified 23,622 proteoforms 2332 proteins from two lines, representing nearly fivefold improvement in number identifications (IDs) compared to previous TDP datasets cells. revealed substantial...

Top-down liquid chromatography-mass spectrometry (LC-MS) analyzes intact proteoforms and generates mass spectra containing peaks of with various isotopic compositions, charge states, retention times. An essential step in top-down MS data analysis is proteoform feature detection, which aims to group these into peak sets (features), each all a proteoform. Accurate protein detection enhances the accuracy MS-based identification quantification. Here, we present TopFD, software tool for that...

Cadmium (Cd) pollution seriously reduces the yield and quality of vegetables. Reducing Cd accumulation in vegetables is great significance for improving food safety sustainable agricultural development. Here, using tomato as material, we analyzed effect foliar spraying with zinc oxide nanoparticles (ZnO NPs) on tolerance seedlings. Foliar ZnO NPs improved by increasing photosynthesis efficiency antioxidative capacity, while it reduced 40.2% roots 34.5% leaves but increased Zn content 33.9%...

A new type of IMAC material, with ATP as the chelating ligand, was synthesized and applied to capture phosphopeptides. For first time, approach for phosphopeptide enrichment could provide selectivity under 5000-fold dilution by nonphosphopeptides, sensitivity on-target at 3 amol.

Abstract Capillary zone electrophoresis (CZE)–tandem mass spectrometry (MS/MS) has recently attracted attention as a tool for shotgun proteomics. However, its performance this analysis so far fallen below that of reversed‐phase liquid chromatography (RPLC)–MS/MS. The use CZE method with wide separation window (up to 90 min) and high peak capacity (ca. 300) is reported. This was coupled an Orbitrap Fusion spectrometer through electrokinetically pumped sheath‐flow interface the complex...

Capillary zone electrophoresis-electrospray ionization-tandem mass spectrometry (CZE-ESI-MS/MS) has been recognized as an invaluable platform for top-down proteomics. However, the scale of proteomics using CZE-MS/MS is still limited due to low loading capacity and narrow separation window CZE. In this work, first time we systematically evaluated dynamic pH junction method focusing intact proteins during CZE-MS. The optimized junction-based approached a 1 μL capacity, 90 min window, high peak...

We report the use of dynamic pH junction based capillary zone electrophoresis-electrospray ionization-tandem mass spectrometry (CZE-ESI-MS/MS) for bottom-up proteomics with an electrokinetically pumped sheath-flow nanospray electrophoresis-mass (CE-MS) interface and both LTQ-XL LTQ-Orbitrap-Velos spectrometers. Conventional injection 20 nL a 1 mg/mL BSA digest identified 37 peptides produced 66% sequence coverage. In contrast, 130 (or larger) 0.05 40 70% coverage using 6.5 sample buffer LTQ....

Native proteomics aims to characterize complex proteomes under native conditions and ultimately produces a full picture of endogenous protein complexes in cells. It requires novel analytical platforms for high-resolution liquid-phase separation prior mass spectrometry (MS) MS/MS. In this work, size-exclusion chromatography (SEC)-capillary zone electrophoresis (CZE)-MS/MS was developed discovery mode, resulting the identification 144 proteins, 672 proteoforms, 23 from Escherichia coli...

Better peptide separation is required for bottom-up proteomics further improving the proteome coverage. The two-dimensional liquid chromatography (2D-LC) systems only explore differences among peptides in their hydrophobicity (reversed-phase, RP) and charge (strong cation/anion exchange, SCX/SAX). Alternative techniques with different mechanisms are to improve separation. Capillary zone electrophoresis (CZE) an attractive alternative because it has high efficiency of biomolecules separates...