Online Parallel Accumulation–Serial Fragmentation (PASEF) with a Novel Trapped Ion Mobility Mass Spectrometer

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Highlights

• •Online PASEF achieves greater than 100 MS/MS per second at full sensitivity.
• •Accurate label-free quantification of over 6000 proteins in 2 h.
• •High throughput demonstrated on 50 ng digests measured in 5 min.
• •High-precision determination of 100,000 peptide collisional cross sections.

     

Comprehensive Analysis of Individual Variation in the Urinary Proteome Revealed Significant Gender Differences

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Highlights

• •Comprehensive analysis of inter-individual variation of normal urinary proteome.
• •Significant gender differences were observed.
• •Proteins increased in female urine are enriched in immunological pathways.
• •Estimated reference intervals of proteins as the baseline for biomarker discovery.

     

Development of a Gill Assay Library for Ecological Proteomics of Threespine Sticklebacks (Gasterosteus aculeatus)

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Highlights

• •Construction of threespine stickleback gill assay library using DDA proteomics
• •Population-specific gill proteome signatures of four ecotypes identified by DIA
• •HSP47 and extracellular matrix proteins highly elevated in warm-adapted sticklebacks
• •Inflammasome and proteolytic proteins highly elevated in freshwater sticklebacks

     

HLA-DO Modulates the Diversity of the MHC-II Self-peptidome

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Highlights

• •MHC-II-bound peptide repertoires from DO-sufficient and DO-deficient cells.
• •Fewer unique peptides and core epitopes were presented in the absence of DO.
• •Immunopeptidome differences appeared to result from reduced DM editing.
• •DO-dependent self-epitopes elicited CD4 T cell responses in mice.

     

Spatiotemporal Changes of the Phagosomal Proteome in Dendritic Cells in Response to LPS Stimulation*

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Highlights

• •Characterization of the phagosomal proteome comparing resting and LPS-treated BMDCs.
• •Label-free quantification determined 2843 phagosomal proteins.
• •Reduced recruitment of hydrolases and V-ATPase to phagosomes of LPS-treated cells.
• •Increased recruitment of antigen cross-presentation molecules to these phagosomes.

     

NNAlign_MA; MHC Peptidome Deconvolution for Accurate MHC Binding Motif Characterization and Improved T-cell Epitope Predictions

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Highlights

• •NNAlign_MA enables full deconvolution of single MHC specificities from MS assays.
• •NNAlign_MA expands MHC allelic coverage, improving identification of T-cell epitopes.
• •NNAlign_MA was benchmarked on MHC classes I and II, outperforming current methods.
• •NNAlign_MA offers a universal solution to analyze and exploit MHC peptidomics data.

     

Integrated Identification and Quantification Error Probabilities for Shotgun Proteomics

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Highlights

• •Unified identification and quantification error rates for protein quantification.
• •Error propagation using graphical models and Bayesian statistics.
• •Account for uncertainty of missing values instead of overconfident point estimates.
• •Control of differential expression false discovery rate at increased sensitivity.

     

Reductive Stress Selectively Disrupts Collagen Homeostasis and Modifies Growth Factor-independent Signaling Through the MAPK/Akt Pathway in Human Dermal Fibroblasts

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Highlights

• •Changes to the proteome of skin fibroblasts subjected to reductive stress have been quantitated.
• •Only a small set of proteins is selectively diminished upon exposure to reductants.
• •Collagens (COL1A2 and COL6A2) emerge as sentinels of reductive stress.
• •Reductive stress triggers receptor-independent Akt phosphorylation at Ser473.

     

Absolute Quantification of Apolipoproteins Following Treatment with Omega-3 Carboxylic Acids and Fenofibrate Using a High Precision Stable Isotope-labeled Recombinant Protein Fragments Based SRM Assay

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Highlights

• •The first application of stable isotope-labeled recombinant protein fragments internal standards (SIS PrEST) on clinical samples.
• •Development of a high-precision quantitative SIS PrEST based LC-SRM Tier 2 assay for 13 apolipoproteins.
• •Semi-automated workflow enables precise and robust high-throughput sample processing.
• •The assay quantifies the effects of feNofibrate and omega-3 carboxylic acids on apolipoproteins in human plasma.

     

Data-independent Acquisition Improves Quantitative Cross-linking Mass Spectrometry

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Highlights

• •Quantitative cross-linking mass spectrometry (QCLMS) was automated by Spectronaut.
• •Data-independent acquisition (DIA) was adapted to QCLMS.
• •Accuracy and precision of quantitation improves with DIA over DDA.
• •QCLMS is now ready for use in complex samples.

     

Analysis of 1508 Plasma Samples by Capillary-Flow Data-Independent Acquisition Profiles Proteomics of Weight Loss and Maintenance

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Highlights

• •Robust capillary-flow DIA was established at 31 clinical samples per day.
• •1508 samples of dietary intervention study DiOGenes were measured on a single column.
• •Comprehensive biological reactions to weight loss and maintenance were observed.
• •Comparison to independent studies shows high reproducibility of potential biomarkers.

     

A Quantitative Chemical Proteomic Strategy for Profiling Phosphoprotein Phosphatases from Yeast to Humans

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Highlights

• •Chemical proteomics strategy for quantitative profiling of phosphoprotein phosphatases.
• •Compatible with quantitative multiplexing approaches.
• •Applicable to many samples types including tissues from human to yeast.

     

A Combined N-terminomics and Shotgun Proteomics Approach to Investigate the Responses of Human Cells to Rapamycin and Zinc at the Mitochondrial Level

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Highlights

• •Rapamycin and zinc induce moderate but significant mitochondrial proteome changes.
• •The mitochondrial proteins processing system is robust under subtoxic conditions.
• •Rapamycin and zinc perturb the mitochondrial proteins processing system.
• •Rapamycin and zinc perturb the mitochondrial proteins homeostasis.

     

The Proteome of Pig Spermatozoa Is Remodeled During Ejaculation

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Highlights

• •Proteome of mature boar spermatozoa from cauda epididymal and ejaculated sources were analyzed by iTRAQ-based LC-MS/MS.
• •1,723 sperm proteins identified (974 of Sus scrofa taxonomy).
• •1,602 sperm proteins quantified (960 of Sus scrofa taxonomy).
• •32 Sus scrofa sperm proteins were differentially expressed among sperm sources.
• •The proteome of boar spermatozoa is remodelled during ejaculation.

     

Simultaneous Improvement in the Precision,Accuracy, and Robustness of Label-free Proteome Quantification by Optimizing Data Manipulation Chains,

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Highlights

• •High-quality LFQ is valuable technique yet remains extremely challenging.
• •Fluctuating precision, limited robustness, and compromised accuracy are known issues.
• •We proposed a strategy collectively improving LFQ precision, robustness, and accuracy.
• •An online tool incorporating this novel strategy was also developed.

     

Intact Transition Epitope Mapping – Targeted High-Energy Rupture of Extracted Epitopes (ITEM-THREE),

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Highlights

• •Multiplex epitope mapping/antigenic determinant identification in the gas phase.
• •Intact transition and controlled dissociation of immune complexes by MS.
• •Simultaneous identification and amino acid sequence determination of epitopes.
• •Simplified in-solution sample handling because of ion manipulation and filtering by MS.

     

Simultaneous Quantification of Protein Expression and Modifications by Top-down Targeted Proteomics: A Case of the Sarcomeric Subproteome

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Highlights

• •A new strategy for simultaneous quantification of protein expression and modification.
• •This top-down LC/MS-based method shows high reproducibility and high throughput.
• •Quantification at the intact protein level with results comparable to Western blot.
• •This top-down proteomics method is applicable to different species and tissues.

     

An Advanced Strategy for Comprehensive Profiling of ADP-ribosylation Sites Using Mass Spectrometry-based Proteomics*

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Highlights

• •Comparing proteolytic digestions and precursor fragmentation methods for MS of ADPr
• •Identification of 11,265 unique ADPr-modified peptides
• •Mapping of hundreds of peptides co-modified by phosphorylation and ADPr
• •ADPr modification of specific residue types displays spatial preferences

     

Comprehensive Analysis of the Lysine Succinylome and Protein Co-modifications in Developing Rice Seeds

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Highlights

• •Nonenzymatically Ksu proteins shown different pattern from native cell Ksu proteins.
• •Motif preference of Ksu proteins was associated with different biological processes.
• •Up to 67 developing rice seeds proteins contain PTMs of Kac, Ksu, Kcr, Kmal, and Khib.
• •Some lysine residues of the key pathway enzymes are modified by succinylation.