Subcellular Peptide Localization in Single Identified Neurons by Capillary Microsampling Mass Spectrometry.
Subcellular Peptide Localization in Single Identified Neurons by Capillary Microsampling Mass Spectrometry.
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DOI：
10.1038/s41598-018-29704-z
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发表时间：
2018-08-15
影响因子：
4.6
通讯作者：
Vertes A
中科院分区：
文献类型：
--
作者：
Zhang L;Khattar N;Kemenes I;Kemenes G;Zrinyi Z;Pirger Z;Vertes A
单细胞质谱法（MS）在罕见细胞中肽的测序和鉴定方面具有独特的定位。小肽可以在亚细胞区室中发挥不同的作用。一些多肽在细胞质中充当神经递质，它们也可以在细胞核中充当转录因子。因此，有必要分析鉴定单细胞中的亚细胞肽组成。在这里，我们应用毛细管微采样质谱和离子迁移分离技术对软体动物停滞淋巴（Lymnaea滞动）单个神经元中的肽进行测序，并对已知表达具有心脏活性的Phe-Met-Arg-Phe酰胺样（fmrfamide -样）神经肽的单个神经元细胞质和细胞核之间的肽分布进行分析。对1型和2型F组（Fgp）神经元的细胞核和细胞质进行了分析，以寻找由FMRFamide基因的选择性剪接产物编码的蛋白前体切割的神经肽。测定了9种神经肽在细胞质中的相对丰度。细胞核中含有六种不同丰度的多肽。利用其在Fgp神经元中的相对富集，用串联质谱法对一个新的28残基神经肽进行了测序。
Single cell mass spectrometry (MS) is uniquely positioned for the sequencing and identification of peptides in rare cells. Small peptides can take on different roles in subcellular compartments. Whereas some peptides serve as neurotransmitters in the cytoplasm, they can also function as transcription factors in the nucleus. Thus, there is a need to analyze the subcellular peptide compositions in identified single cells. Here, we apply capillary microsampling MS with ion mobility separation for the sequencing of peptides in single neurons of the mollusk Lymnaea stagnalis, and the analysis of peptide distributions between the cytoplasm and nucleus of identified single neurons that are known to express cardioactive Phe-Met-Arg-Phe amide-like (FMRFamide-like) neuropeptides. Nuclei and cytoplasm of Type 1 and Type 2 F group (Fgp) neurons were analyzed for neuropeptides cleaved from the protein precursors encoded by alternative splicing products of the FMRFamide gene. Relative abundances of nine neuropeptides were determined in the cytoplasm. The nuclei contained six of these peptides at different abundances. Enabled by its relative enrichment in Fgp neurons, a new 28-residue neuropeptide was sequenced by tandem MS.
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影响因子：
48
作者：
Giesen, Charlotte;Wang, Hao A. O.;Bodenmiller, Bernd
通讯作者：
Bodenmiller, Bernd
影响因子：
5.3
作者：
BRIGHT, K;KELLETT, E;BENJAMIN, PR
通讯作者：
BENJAMIN, PR
影响因子：
3.4
作者：
Janes, Tara A.;Xu, Fenglian;Syed, Naweed I.
通讯作者：
Syed, Naweed I.
影响因子：
3.7
作者：
Burns, Tyler J.;Frei, Andreas P.;Nolan, Garry P.
通讯作者：
Nolan, Garry P.
影响因子：
64.5
作者：
Chen, Walter W.;Freinkman, Elizaveta;Wang, Tim;Birsoy, Kivanc;Sabatini, David M.
通讯作者：
Sabatini, David M.