8 research outputs found
Genomics and proteomics in chemical warfare agent research: Recent studies and future applications
A Large-Scale Quantitative Proteomic Approach To Identifying Sulfur Mustard-Induced Protein Phosphorylation Cascades
Combining Amine Metabolomics and Quantitative Proteomics of Cancer Cells Using Derivatization with Isobaric Tags
Quantitative metabolomics and proteomics
technologies are powerful
approaches to explore cellular metabolic regulation. Unfortunately,
combining the two technologies typically requires different LC-MS
setups for sensitive measurement of metabolites and peptides. One
approach to enhance the analysis of certain classes of metabolites
is by derivatization with various types of tags to increase ionization
and chromatographic efficiency. We demonstrate here that derivatization
of amine metabolites with tandem mass tags (TMT), typically used in
multiplexed peptide quantitation, facilitates amino acid analysis
by standard nanoflow reversed-phase LC-MS setups used for proteomics.
We demonstrate that this approach offers the potential to perform
experiments at the MS1-level using duplex tags or at the MS2-level
using novel 10-plex reporter ion-containing isobaric tags for multiplexed
amine metabolite analysis. We also demonstrate absolute quantitative
measurements of amino acids conducted in parallel with multiplexed
quantitative proteomics, using similar LC-MS setups to explore cellular
amino acid regulation. We further show that the approach can also
be used to determine intracellular metabolic labeling of amino acids
from glucose carbons
Combining Amine Metabolomics and Quantitative Proteomics of Cancer Cells Using Derivatization with Isobaric Tags
Quantitative metabolomics and proteomics
technologies are powerful
approaches to explore cellular metabolic regulation. Unfortunately,
combining the two technologies typically requires different LC-MS
setups for sensitive measurement of metabolites and peptides. One
approach to enhance the analysis of certain classes of metabolites
is by derivatization with various types of tags to increase ionization
and chromatographic efficiency. We demonstrate here that derivatization
of amine metabolites with tandem mass tags (TMT), typically used in
multiplexed peptide quantitation, facilitates amino acid analysis
by standard nanoflow reversed-phase LC-MS setups used for proteomics.
We demonstrate that this approach offers the potential to perform
experiments at the MS1-level using duplex tags or at the MS2-level
using novel 10-plex reporter ion-containing isobaric tags for multiplexed
amine metabolite analysis. We also demonstrate absolute quantitative
measurements of amino acids conducted in parallel with multiplexed
quantitative proteomics, using similar LC-MS setups to explore cellular
amino acid regulation. We further show that the approach can also
be used to determine intracellular metabolic labeling of amino acids
from glucose carbons
Rapid, deep and precise profiling of the plasma proteome with multi-nanoparticle protein corona
© 2020, The Author(s). Large-scale, unbiased proteomics studies are constrained by the complexity of the plasma proteome. Here we report a highly parallel protein quantitation platform integrating nanoparticle (NP) protein coronas with liquid chromatography-mass spectrometry for efficient proteomic profiling. A protein corona is a protein layer adsorbed onto NPs upon contact with biofluids. Varying the physicochemical properties of engineered NPs translates to distinct protein corona patterns enabling differential and reproducible interrogation of biological samples, including deep sampling of the plasma proteome. Spike experiments confirm a linear signal response. The median coefficient of variation was 22%. We screened 43 NPs and selected a panel of 5, which detect more than 2,000 proteins from 141 plasma samples using a 96-well automated workflow in a pilot non-small cell lung cancer classification study. Our streamlined workflow combines depth of coverage and throughput with precise quantification based on unique interactions between proteins and NPs engineered for deep and scalable quantitative proteomic studies