Label-Free Quantitative Proteomics Reveals Differentially Regulated Proteins in Experimental Gingivitis
Label-Free Quantitative Proteomics Reveals Differentially Regulated Proteins in Experimental Gingivitis
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DOI：
10.1021/pr300761e
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发表时间：
2013-02-01
影响因子：
4.4
通讯作者：
Papapanou, Panos N.
中科院分区：
文献类型：
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作者：
Bostanci, Nagihan;Ramberg, Per;Papapanou, Panos N.
我们研究了在实验性牙龈炎的诱导（I）和消退（R）过程中龈沟液样品中的顺序蛋白质表达。牙周和全身健康的志愿者（n = 20）参加了为期三周的实验牙龈炎协议，清创和两个星期的定期菌斑控制。在基线、第7、14和21天（诱导期; I期）以及第21、25、30和35天（消退期; R期）采集龈沟液（GCF）样本。液相色谱串联质谱（LC-MS/MS）用于无标记定量蛋白质组学。共鉴定了287种蛋白质，包括254种人类蛋白质、14种细菌蛋白质、12种真菌蛋白质和7种酵母蛋白质。本体论分析显示蛋白质主要参与细胞骨架重排、免疫反应、抗菌功能、蛋白质降解和DNA结合。在不同时间点，受试者之间和受试者内鉴定的蛋白质数量存在相当大的差异。在每个时间点合并受试者之间的样品后，纵向定量I期中的59种蛋白质和R期中的73种蛋白质的水平。我们的数据表明，LC MS/MS无标记定量蛋白质组学在评估GCF的蛋白质含量方面是有价值的，并且可以促进更好地理解参与人类牙菌斑诱导的牙龈炎症的诱导和解决的分子机制。
We investigated the sequential protein expression in gingival crevicular fluid samples during the induction (I) and resolution (R) of experimental gingivitis. Periodontally and systemically healthy volunteers (n = 20) participated in a three-week experimental gingivitis protocol, followed by debridement and two weeks of regular plaque control. Gingival crevicular fluid (GCF) samples were collected at baseline, Day 7, 14, and 21 (induction; I-phase), and at Day 21, 25, 30, and 35 (resolution; R-phase). Liquid chromatography tandem mass spectrometry (LC-MS/MS) for label-free quantitative proteomics was applied. A total of 287 proteins were identified including 254 human, 14 bacterial, 12 fungal, and 7 yeast proteins. Ontology analysis revealed proteins primarily involved in cytoskeletal rearrangements, immune response, antimicrobial function, protein degradation, and DNA binding. There was considerable variation in the number of proteins identified, both among subjects and within subjects across time points. After pooling of samples between subjects at each time point, the levels of 59 proteins in the I-phase and 73 proteins in the R-phase were quantified longitudinally. Our data demonstrate that LC MS/MS label-free quantitative proteomics is valuable in the assessment of the protein content of the GCF and can facilitate a better understanding of the molecular mechanisms involved in the induction and resolution of plaque-induced gingival inflammation in humans.