Analysis of Peptide Stereochemistry in Single Cells by Capillary Electrophoresis–Trapped Ion Mobility Spectrometry Mass Spectrometry
Autor: | Jonathan V. Sweedler, Hsiao Wei Liao, David H. Mast, Elena V. Romanova |
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Rok vydání: | 2021 |
Předmět: |
Proteomics
chemistry.chemical_classification Neuropeptide Gene Ion-mobility spectrometry Stereochemistry 010401 analytical chemistry Cell Electrophoresis Capillary Peptide 010402 general chemistry Mass spectrometry 01 natural sciences Mass Spectrometry Article 0104 chemical sciences Analytical Chemistry medicine.anatomical_structure Capillary electrophoresis Ion-mobility spectrometry–mass spectrometry chemistry Single-cell analysis Ion Mobility Spectrometry medicine Peptides |
Zdroj: | Anal Chem |
ISSN: | 1520-6882 0003-2700 |
Popis: | Single cell analysis strives to probe molecular heterogeneity in morphologically similar cell populations through quantitative or qualitative measurements of genetic, proteomic, or metabolic products. Here, we applied mass analysis of single neurons to investigate cell-cell signaling peptides. The multiplicity of endogenous cell-cell signaling peptides is a common source of chemical diversity among cell populations. Certain peptides can undergo post-translational isomerization of select residues, which has important physiological consequences. The limited number of single cell analysis techniques that are sensitive to peptide stereochemistry make it challenging to study isomerization at the individual cell level. We performed capillary electrophoresis (CE) with mass spectrometry (MS) detection to characterize the peptide content of single cells. Using complementary trapped ion mobility spectrometry (TIMS) separations, we measured the stereochemical configurations of three neuropeptide gene products derived from the pleurin precursor in individual neurons (N = 3) isolated from the central nervous system of Aplysia californica. An analysis of the resultant mobility profiles indicated >98% of the detectable pleurin-derived peptides exist as the non-isomerized, all-L forms in individual neuron cell bodies. However, we observed 44% of the Plrn2 peptide from the pleurin precursor was present as the isomerized, D-residue-containing form in the nerve tissue. These findings demonstrate an unusual distribution of isomerized peptides in A. californica and establish CE–TIMS MS as a powerful analytical tool for investigating peptide stereochemistry at the single cell level. |
Databáze: | OpenAIRE |
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