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首页> 外文期刊>International Journal of Peptide Research and Therapeutics >Mass-Spectrometry Based Characterisation of Infant Whole Saliva Peptidome
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Mass-Spectrometry Based Characterisation of Infant Whole Saliva Peptidome

机译：基于质谱的婴儿全唾液肽组表征

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摘要

The objective of the study was to determine optimal conditions for sampling, sample processing and mass-spectrometry based analysis of infants’ salivary peptidome. Saliva was sampled in 3- and 6-month-old infants and peptide extracts were prepared. Various sample pretreatments before profiling by MALDI-ToF were evaluated and peptide identification was undertaken by MALDI-ToF/ToF or nanoLC-ESI-IT tandem MS. A fast and simple protocol (cut-off filtration at 5 kDa) was satisfactory to produce extracts where no proteolysis was detected even when no protease inhibitor was added. Optimal MALDI spectra were generated after purification on C18 tips. Variability of spectra between two samples exceeded that of the technical replicates, validating that the method is suitable to conduct differential studies. Salivary peptides, identified by means of the two complementary mass spectrometry techniques, were fragments of proline-rich proteins and histatins. The fragments originated mainly from the C-terminal protein extremities. Indications on the proteolytic systems involved and the anatomic location where they intervene are proposed.
机译：该研究的目的是确定婴儿唾液肽组分析的最佳条件，以进行采样，样品处理和质谱分析。在3个月和6个月大的婴儿中取样唾液，并制备了肽提取物。在通过MALDI-ToF进行分析之前，对各种样品预处理进行了评估，并通过MALDI-ToF / ToF或nanoLC-ESI-IT串联MS对肽进行了鉴定。快速简便的操作流程（在5 kDa处进行截止过滤）足以产生即使未添加蛋白酶抑制剂也未检测到蛋白水解的提取物。在C18吸头上纯化后产生最佳MALDI光谱。两个样品之间的光谱变异性超过了技术复制的光谱变异性，这证明该方法适合进行差异研究。通过两种互补质谱技术鉴定的唾液肽是富含脯氨酸的蛋白质和组蛋白的片段。片段主要起源于C末端蛋白末端。提出了有关所涉及的蛋白水解系统以及它们所介入的解剖位置的指示。

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  • 作者

    G. Lucchi; C. Chambon; C. Truntzer; D. Pecqueur; P. Ducoroy; C. Schwartz; S. Nicklaus; M. Morzel;

  • 作者单位

    CLIPP « Clinical Innovation Proteomic Platform » IFR 100 Santé-STIC Université de Bourgogne CHU Le Bocage 21000 Dijon France;

    INRA Plate-forme « Exploration du métabolisme: des gènes aux métabolites » 63122 St Genès Champanelle France;

    CLIPP « Clinical Innovation Proteomic Platform » IFR 100 Santé-STIC Université de Bourgogne CHU Le Bocage 21000 Dijon France;

    CLIPP « Clinical Innovation Proteomic Platform » IFR 100 Santé-STIC Université de Bourgogne CHU Le Bocage 21000 Dijon France;

    CLIPP « Clinical Innovation Proteomic Platform » IFR 100 Santé-STIC Université de Bourgogne CHU Le Bocage 21000 Dijon France;

    INRA UMR 1129 FLAVIC 17 rue Sully 21000 Dijon France;

    INRA UMR 1129 FLAVIC 17 rue Sully 21000 Dijon France;

    INRA UMR 1129 FLAVIC 17 rue Sully 21000 Dijon France;

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  • 原文格式 PDF
  • 正文语种 eng
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  • 关键词

    Saliva peptidome; Infants; Mass spectrometry; MALDI-ToF; Sample pretreatment; PRPs;

    机译：唾液肽组;婴儿;质谱;MALDI-ToF;样品预处理;PRPs;

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