Raw data are available via the MassIVE repository with identifier MSV000087476.
#Theofil look wisconsin pro
Overall, we anticipate this ultrafast, robust, and reproducible bottom-up method empowered by both Azo and the timsTOF Pro will be generally applicable and greatly accelerate the throughput of large-scale quantitative proteomic studies. We applied this method to analyze the complex human cardiac proteome and identified nearly 4000 protein groups from as little as 1 mg of human heart tissue in a single one-dimensional LC-TIMS-MS/MS run with high reproducibility. Herein, we developed an ultrafast bottom-up proteomics method enabled by Azo, a photocleavable, MS-compatible surfactant that effectively solubilizes proteins and promotes rapid tryptic digestion, combined with the Bruker timsTOF Pro, which enables deeper proteome coverage through trapped ion mobility spectrometry (TIMS) and parallel accumulation-serial fragmentation (PASEF) of peptides. Thus, there is a need for a fast, robust, and reproducible method for protein identification and quantification from complex proteomes.
#Theofil look wisconsin Offline
However, traditional sample preparation methods are time-consuming, typically including overnight tryptic digestion, extensive sample cleanup to remove MS-incompatible surfactants, and offline sample fractionation to reduce proteome complexity prior to online liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. Global bottom-up mass spectrometry (MS)-based proteomics is widely used for protein identification and quantification to achieve a comprehensive understanding of the composition, structure, and function of the proteome. This strategy can be used to generate biological hypotheses and identify biomarker candidates to advance the understanding of hPSC-CM differentiation and maturation. This multi-omics method achieves deep coverage of metabolites and proteins, creating a multidimensional view of the hPSC-CM phenotype. Highly populated pathways included those involved in protein synthesis (ribosome, spliceosome), ATP generation (oxidative phosphorylation), and cardiac muscle contraction. Out of the 310 total pathways identified using metabolomics and proteomics data, 40 pathways were considered significantly overrepresented (FDR-corrected p ≤ 0.05). We further integrated the proteome and metabolome measurements to create network profiles of molecular phenotypes for hPSC-CMs. Using this strategy, we identified an average of 205 metabolites/lipids and 4,008 protein groups from 106 cells with high reproducibility.
Here we developed a sequential extraction to capture metabolites and proteins from hPSC-CM monolayer cultures, and analyzed these extracts using high resolution mass spectrometry (MS). To gain global insight into hPSC-CM biology, we introduce a multi-omics strategy for analyzing the hPSC-CM metabolome and proteome from the same cell culture, creating multi-dimensional profiles of hPSC-CMs. However, hPSC-CMs in culture have not recapitulated the structure or functional properties of adult CMs in vivo thus far. Human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) have shown immense promise for patient-specific disease modeling, cardiotoxicity screening, and regenerative therapy development.
0 kommentar(er)
