Analysis of proteins in human gingival crevicular fluid by mass spectrometry
Analysis of proteins in human gingival crevicular fluid by mass spectrometry
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DOI：
10.1111/j.1600-0765.2011.01458.x
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发表时间：
2012-08-01
影响因子：
3.5
通讯作者：
Nagata, T.
中科院分区：
文献类型：
--
作者：
Kido, J.;Bando, M.;Nagata, T.
Kido J，Bando M，Hiroshima Y，Iwasaka H，Yamada K，Ohgami N，Nambu T，Kataoka M，Yamamoto T，Shinohara Y，Sakawa I，Nagata T。通过质谱法分析人龈沟液中的蛋白质。牙周病研究杂志，2012； 47: 488499. (c) 2012 John Wiley & Sons A/S 背景和目的：龈沟液是从牙周组织渗入龈缝和牙周袋的体液，含有多种成分。龈沟液中的蛋白质已被研究作为牙周疾病的标志物。质谱分析用于蛋白质、脂质、糖类和金属的分析，并有望成为疾病诊断的一种方法。为了更好地分析龈沟液中的蛋白质成分，我们使用质谱法研究了来自健康龈缝和牙周袋的龈沟液样本中的蛋白质。材料和方法：龈沟液样本是从知情同意且牙周健康或牙袋患病的受试者中采集的。通过电泳分离这些样品，并通过纳米液相色谱结合串联质谱分析凝胶上的每个级分。通过蛋白质印迹法证实了龈沟液中检测到的抗菌肽。结果：健康部位和牙周炎部位的龈沟液样本中均检测到104种蛋白质； 64 种蛋白质仅在健康部位的龈沟液中含有，63 种蛋白质仅在牙周炎部位的龈沟液中观察到。这些蛋白质是与血液、细胞骨架、免疫、炎症和脂质相关的蛋白质和酶。首次在龈沟液中鉴定出铜蓝蛋白、糖原磷酸化酶、谷胱甘肽S-转移酶、磷酸甘油酸变位酶、银屑病素、S100A11和抵抗素等蛋白质。通过质谱和蛋白质印迹法观察抗菌肽，如乳铁蛋白、α1-抗胰蛋白酶、脂质运载蛋白、S100A7、S100A8、S100A9 和抗菌肽。结论：采用质谱技术同时分析龈沟液中的多种蛋白质成分，该方法可能有助于牙周病的诊断。
Kido J, Bando M, Hiroshima Y, Iwasaka H, Yamada K, Ohgami N, Nambu T, Kataoka M, Yamamoto T, Shinohara Y, Sagawa I, Nagata T. Analysis of proteins in human gingival crevicular fluid by mass spectrometry. J Periodont Res 2012; 47: 488499. (c) 2012 John Wiley & Sons A/S Background and Objective: Gingival crevicular fluid is a bodily fluid transuded from periodontal tissues into the gingival crevice and periodontal pocket, and contains many species of components. Proteins in gingival crevicular fluid have been studied as markers for periodontal diseases. Mass spectrometric analysis is used for the analyses of proteins, lipids, saccharides and metals, and expected as an approach for disease diagnosis. For better analysis of the protein components in gingival crevicular fluid, we investigated proteins in gingival crevicular fluid samples from the healthy gingival crevice and periodontal pocket using mass spectrometry. Material and Methods: Gingival crevicular fluid samples were collected from subjects who gave their informed consent and were periodontally healthy or had diseased pockets. These samples were electrophoretically separated, and each fraction on the gels was analysed by nano liquid chromatography coupled with tandem mass spectrometry. Antimicrobial peptides detected in gingival crevicular fluid were confirmed by western blotting. Results: One hundred and four proteins were detected in gingival crevicular fluid samples from both healthy sites and sites of periodontitis; 64 proteins were contained only in gingival crevicular fluid from healthy sites and 63 proteins were observed only in gingival crevicular fluid from periodontitis sites. These proteins were blood-, cytoskeleton-, immunity-, inflammation- and lipid-related proteins and enzymes. Some proteins, including ceruloplasmin, glycogen phosphorylase, glutathione S-transferase, phosphoglycerate mutase, psoriasin, S100A11 and resistin, were identified for the first time in gingival crevicular fluid. Antimicrobial peptides, such as lactoferrin, a1-antitrypsin, lipocalin, S100A7, S100A8, S100A9 and cathelicidin, were observed by mass spectrometry and western blotting. Conclusion: Multiple protein components in gingival crevicular fluid were analysed at the same time using mass spectrometry, and this approach may be useful for the diagnosis of periodontal diseases.