Analysis of Peptide Stereochemistry in Single Cells by Capillary Electrophoresis-Trapped Ion Mobility Spectrometry Mass Spectrometry.
Analysis of Peptide Stereochemistry in Single Cells by Capillary Electrophoresis-Trapped Ion Mobility Spectrometry Mass Spectrometry.
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DOI：
10.1021/acs.analchem.1c00445
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发表时间：
2021-04-20
影响因子：
7.4
通讯作者：
Sweedler JV
中科院分区：
文献类型：
--
作者：
Mast DH;Liao HW;Romanova EV;Sweedler JV
单细胞分析致力于通过对遗传、蛋白质组或代谢产物的定量或定性测量来探索形态相似的细胞群体中的分子异质性。在这里，我们应用单个神经元的质量分析来研究细胞-细胞信号肽。内源性细胞-细胞信号肽的多样性是细胞群体间化学多样性的共同来源。某些多肽可以发生选择性残基的翻译后异构化，这具有重要的生理后果。对多肽立体化学敏感的单细胞分析技术的数量有限，这使得在单个细胞水平上研究异构化具有挑战性。用毛细管电泳法(CE)和质谱仪(MS)检测单细胞的多肽含量。利用互补俘获离子迁移率谱(TIMS)分离技术，我们测定了从海兔中枢神经系统分离的单个神经元(N=3)中三种神经肽基因产物的立体化学构型。对所得迁移率分布的分析表明，98%可检测到的侧柏醇衍生物多肽以非异构化的全L形式存在于单个神经元细胞体中。然而，我们观察到来自胸苷前体的Plrn2多肽中有44%是以异构化的、含有D残基的形式存在于神经组织中的。这些发现证明了异构化多肽在管胞内的异常分布，并建立了CE-TIMS MS作为在单细胞水平上研究多肽立体化学的强大分析工具。
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.
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影响因子：
4
作者：
Mast, David H.;Checco, James W.;Sweedler, Jonathan V.
通讯作者：
Sweedler, Jonathan V.
影响因子：
17.3
作者：
Li, LJ;Garden, RW;Sweedler, JV
通讯作者：
Sweedler, JV
DOI：
10.1039/c2an16211c
发表时间：
2012-07-07
期刊：
The Analyst
影响因子：
--
作者：
Cecala C;Sweedler JV
通讯作者：
Sweedler JV
影响因子：
4.3
作者：
Lombard-Banek, Camille;Yu, Zhe;Nemes, Peter
通讯作者：
Nemes, Peter
影响因子：
7.4
作者：
Adams, CM;Zubarev, RA
通讯作者：
Zubarev, RA