A new albumin-depletion strategy improves proteomic research of gingival crevicular fluid from periodontitis patients
A new albumin-depletion strategy improves proteomic research of gingival crevicular fluid from periodontitis patients
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DOI：
10.1007/s00784-017-2213-0
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发表时间：
2018-04-01
影响因子：
3.4
通讯作者：
Lenz, Christof
中科院分区：
文献类型：
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作者：
Batschkus, Sarah;Cingoez, Goekhan;Lenz, Christof
龈沟液（Gingival crevicular fluid，GCF）是龈沟内的炎性浸润物，在口腔的各种过程中具有重要意义，在口腔科学中具有重要的影响。它被认为是系统性或牙周疾病的生物标志物和正畸牙齿移动的介质的来源。为了以无偏和完整的方法表征GCF的蛋白质含量，我们采用了质谱法（MS），其不仅允许鉴定，而且还可以定量这些蛋白质。在从患有牙周炎的患者获得的样品中，由于存在源自血清的高度丰富的血清白蛋白，该方法通常受到限制。本研究的目的是采用蛋白质沉淀程序有效去除GCF样本中的血清白蛋白。从五名成人牙周炎患者中收集的GFC样本通过三氯乙酸/丙酮沉淀进行分级，并通过SDS-PAGE和高分辨率质谱分析所得的可溶性和沉淀部分。三氯乙酸/丙酮沉淀被成功地用作从GCF样品中有效去除血清白蛋白的蛋白质沉淀程序。仔细分析发现，沉淀步骤有效地降低了血清白蛋白含量，并将随后的蛋白质鉴定提高了32%。新的蛋白质组学方法可以鉴定出317个蛋白质，GCF蛋白质组覆盖率的增加将有助于我们更好地了解牙周组织在牙周炎发病过程中的分子机制，我们新的白蛋白去除策略结合高分辨质谱技术可以有效地监测牙周组织的分子信号。
Gingival crevicular fluid (GCF), the inflammatory infiltrate within the crevicular sulcus, is of great importance for diverse processes in the oral cavity and has a high impact in oral sciences. It is assumed to serve as a source of biomarkers for systemic or periodontal diseases and mediators of orthodontic tooth movement. In order to characterize the protein content of the GCF in an unbiased and complete approach, we employed mass spectrometry (MS), which allows not only the identification, but also the quantification of these proteins. In samples obtained from patients suffering from periodontitis, this method is often limited due to the presence of highly abundant serum albumin deriving from serum. The aim of this investigation was to employ a protein precipitation procedure for the efficient depletion of serum albumin from GCF samples.GFC samples collected from five adult periodontitis patients were fractionated by trichloroacetic acid/acetone precipitation and the resulting soluble and pelleted fractions were analyzed by SDS-PAGE and high-resolution mass spectrometry.Trichloroacetic acid/acetone precipitation was successfully employed as a protein precipitation procedure for the efficient depletion of serum albumin from GCF samples. Careful analysis revealed that the precipitation step reduced the serum albumin content efficiently, and increased subsequent protein identifications by 32%. Three hundred seventeen proteins could only be identified with this new approach.The increased coverage of the GCF proteome will help improve our understanding of molecular mechanisms in the periodontium during pathogenesis of periodontitis.Our new albumin depletion strategy combined with high-resolution mass spectrometry can be used to effectively monitor the molecular signals of the periodontium.