iTRAQ-based quantitative analysis of age-specific variations in salivary proteome of caries-susceptible individuals.
iTRAQ-based quantitative analysis of age-specific variations in salivary proteome of caries-susceptible individuals.
复制标题
基于 iTRAQ 的龋齿易感个体唾液蛋白质组年龄特异性变异定量分析
DOI：
10.1186/s12967-018-1669-2
复制
发表时间：
2018-10-25
影响因子：
7.4
通讯作者：
Zhang L
中科院分区：
文献类型：
--
作者：
Wang K;Wang X;Zheng S;Niu Y;Zheng W;Qin X;Li Z;Luo J;Jiang W;Zhou X;Li W;Zhang L
背景人类唾液是一种富含蛋白质、易于获取的潜在生物标志物来源，可用于诊断口腔和全身疾病。然而，人们对与龋齿患者衰老相关的唾液蛋白质组变化知之甚少。在这里，我们应用同量异位标签进行相对和绝对定量（iTRAQ）结合多反应监测质谱（MRM-MS）来表征不同年龄受试者（有龋齿和无龋齿）的唾液蛋白质组图谱，旨在识别与年龄相关的龋齿生物标志物。方法从40名无龋齿和易患龋齿的年轻人和老年人中收集未刺激的全唾液样本。唾液蛋白被提取、还原、烷基化、胰蛋白酶消化，然后使用 iTRAQ 耦合 LC-MS/MS 进行分析，然后进行 GO 注释、生物途径分析、层次聚类分析和蛋白质-蛋白质相互作用分析。然后使用 MRM-MS 进行候选验证。结果在使用串联质谱法鉴定的 658 个唾液蛋白中，435 个蛋白在有龋齿和无龋齿的不同年龄组中表现出表达模式改变。在这些蛋白质中，96个在易患龋齿的成年人和老年人中表现出年龄特异性变化，并且主要与唾液分泌途径相关，而在健康个体中鉴定出110个年龄特异性蛋白质。研究发现，年龄因素会引起显着的变异，并在健康和致龋唾液蛋白质组中发挥重要作用。随后，使用 MRM 分析进一步成功验证了总共 136 个具有复杂蛋白质-蛋白质相互作用的目标蛋白质，其中包括 14 个与龋齿相关的年龄特异性蛋白质。此外，非年龄特异性蛋白（histatin-1 和 BPI 折叠家族 B 成员 1）被证实是常见龋齿的重要候选生物标志物。结论我们使用发现-验证流程进行的蛋白质组学分析揭示了健康和致龋唾液蛋白质组中由年龄因素引起的明显变化，突出了年龄在唾液用于龋齿诊断和生物标志物发现的巨大潜力中的重要性。
BackgroundHuman saliva is a protein-rich, easily accessible source of potential biomarkers for the diagnosis of oral and systemic diseases. However, little is known about the changes in salivary proteome associated with aging of patients with dental caries. Here, we applied isobaric tags for relative and absolute quantitation (iTRAQ) in combination with multiple reaction monitoring mass spectrometry (MRM-MS) to characterize the salivary proteome profiles of subjects of different ages, presenting with and without caries, with the aim of identifying age-related biomarkers for dental caries.MethodsUnstimulated whole saliva samples were collected from 40 caries-free and caries-susceptible young adults and elderly individuals. Salivary proteins were extracted, reduced, alkylated, digested with trypsin and then analyzed using iTRAQ-coupled LC–MS/MS, followed by GO annotation, biological pathway analysis, hierarchical clustering analysis, and protein–protein interaction analysis. Candidate verification was then conducted using MRM-MS.ResultsAmong 658 salivary proteins identified using tandem mass spectrometry, 435 proteins exhibited altered expression patterns in different age groups with and without caries. Of these proteins, 96 displayed age-specific changes among caries-susceptible adults and elderly individuals, and were mainly associated with salivary secretion pathway, while 110 age-specific proteins were identified among healthy individuals. It was found that the age factor caused significant variations and played an important role in both healthy and cariogenic salivary proteomes. Subsequently, a total of 136 target proteins with complex protein–protein interactions, including 14 age-specific proteins associated with caries, were further successfully validated using MRM analysis. Moreover, non-age-specific proteins (histatin-1 and BPI fold-containing family B member 1) were verified to be important candidate biomarkers for common dental caries.ConclusionsOur proteomic analysis performed using the discovery-through-verification pipeline revealed distinct variations caused by age factor in both healthy and cariogenic salivary proteomes, highlighting the significance of age in the great potential of saliva for caries diagnosis and biomarker discovery.
登录
查看更多内容
DOI：
10.1016/j.matbio.2015.12.007
发表时间：
2016-05
期刊：
Matrix biology : journal of the International Society for Matrix Biology
影响因子：
--
作者：
Duverger O;Beniash E;Morasso MI
通讯作者：
Morasso MI
影响因子：
7.4
作者：
Sun X;Huang X;Tan X;Si Y;Wang X;Chen F;Zheng S
通讯作者：
Zheng S
影响因子：
7.6
作者：
Siqueira, W. L.;Salih, E.;Oppenheim, F. G.
通讯作者：
Oppenheim, F. G.
影响因子：
3.7
作者：
Hong, S. W.;Seo, D. -G.;Han, S. H.
通讯作者：
Han, S. H.
影响因子：
4.5
作者：
Duverger O;Carlson JC;Karacz CM;Schwartz ME;Cross MA;Marazita ML;Shaffer JR;Morasso MI
通讯作者：
Morasso MI