Novel protein identification methods for biomarker discovery via a proteomic analysis of periodontally healthy and diseased gingival crevicular fluid samples.
Novel protein identification methods for biomarker discovery via a proteomic analysis of periodontally healthy and diseased gingival crevicular fluid samples.
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DOI：
10.1111/j.1600-051x.2011.01805.x
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发表时间：
2012-03
影响因子：
6.7
通讯作者：
Floudas CA
中科院分区：
文献类型：
--
作者：
Baliban RC;Sakellari D;Li Z;DiMaggio PA;Garcia BA;Floudas CA
通过高通量蛋白质组分析，识别慢性牙周炎（CP）和牙周健康个体的龈沟液（GCF）样本中可能的新型生物标志物。 GCF 样本采集自 12 名 CP 和 12 名牙周健康受试者。样品经过胰酶消化，使用高效液相色谱进行洗脱，并使用串联质谱 (MS/MS) 进行片段化。使用 PILOT_PROTEIN 进行 MS/MS 分析，以识别样品中所有未修饰的蛋白质。使用源自智人分类学和所有细菌分类学的数据库，鉴定了 432 种人类蛋白（120 种新蛋白）和 30 种细菌蛋白。人类蛋白质、血管紧张素原、簇蛋白和胸苷磷酸化酶仅在牙周健康样本中获得高分，因此被确定为候选生物标志物。同样，中性粒细胞防御素 1、碳酸酐酶 1 和伸长因子 1 γ 也与 CP 相关。候选细菌生物标志物包括 33 kDa 伴侣蛋白、铁摄取蛋白 A2 和磷酸烯醇丙酮酸羧化酶（与健康相关）和核酮糖二磷酸羧化酶、可能的琥珀酰辅酶 A:3-酮酸辅酶 A 转移酶或 DNA 指导的 RNA 聚合酶亚基 β（CP 相关）。大多数这些人类和细菌蛋白质之前尚未被评估为牙周状况的生物标志物，需要进一步研究。所提出的大规模综合蛋白质组分析方法可能有助于鉴定牙周病的新型生物标志物。
To identify possible novel biomarkers in gingival crevicular fluid (GCF) samples from chronic periodontitis (CP) and periodontally healthy individuals by high-throughput proteomic analysis. GCF samples were collected from twelve CP and twelve periodontally healthy subjects. Samples were trypically digested, eluted using high-performance liquid chromatography, and fragmented using tandem mass spectrometry (MS/MS). MS/MS were analyzed using PILOT_PROTEIN to identify all unmodified proteins within the samples. Using the database derived from Homo sapiens taxonomy and all bacterial taxonomies, 432 human (120 new) and 30 bacterial proteins were identified. The human proteins, angiotensinogen, clusterin and thymidine phosphorylase were identified as biomarker candidates based on their high-scoring only in samples from periodontal health. Similarly, neutrophil defensin-1, carbonic anhydrase-1 and elongation factor-1 gamma were associated with CP. Candidate bacterial biomarkers include 33 kDa chaperonin, iron uptake protein A2 and phosphoenolpyruvate carboxylase (health-associated) and ribulose biphosphate carboxylase, a probable succinyl-CoA:3-ketoacid-coenzyme A transferase, or DNA-directed RNA polymerase subunit beta (CP-associated). Most of these human and bacterial proteins have not been previously evaluated as biomarkers of periodontal conditions and require further investigation. The proposed methods for large-scale comprehensive proteomic analysis may lead to the identification of novel biomarkers of periodontal disease.