Belstrøm Daniel, Jersie-Christensen Rosa R, Lyon David, Damgaard Christian, Jensen Lars J, Holmstrup Palle, Olsen Jesper V
Section of Periodontology and Microbiology, Department of Odontology, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
Proteomics Program, Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
PeerJ. 2016 Sep 14;4:e2433. doi: 10.7717/peerj.2433. eCollection 2016.
The composition of the salivary microbiota has been reported to differentiate between patients with periodontitis, dental caries and orally healthy individuals. To identify characteristics of diseased and healthy saliva we thus wanted to compare saliva metaproteomes from patients with periodontitis and dental caries to healthy individuals.
Stimulated saliva samples were collected from 10 patients with periodontitis, 10 patients with dental caries and 10 orally healthy individuals. The proteins in the saliva samples were subjected to denaturing buffer and digested enzymatically with LysC and trypsin. The resulting peptide mixtures were cleaned up by solid-phase extraction and separated online with 2 h gradients by nano-scale C18 reversed-phase chromatography connected to a mass spectrometer through an electrospray source. The eluting peptides were analyzed on a tandem mass spectrometer operated in data-dependent acquisition mode.
We identified a total of 35,664 unique peptides from 4,161 different proteins, of which 1,946 and 2,090 were of bacterial and human origin, respectively. The human protein profiles displayed significant overexpression of the complement system and inflammatory markers in periodontitis and dental caries compared to healthy controls. Bacterial proteome profiles and functional annotation were very similar in health and disease.
Overexpression of proteins related to the complement system and inflammation seems to correlate with oral disease status. Similar bacterial proteomes in healthy and diseased individuals suggests that the salivary microbiota predominantly thrives in a planktonic state expressing no disease-associated characteristics of metabolic activity.
据报道，牙周炎、龋齿患者与口腔健康个体的唾液微生物群组成存在差异。因此，为了确定患病和健康唾液的特征，我们希望比较牙周炎和龋齿患者与健康个体的唾液元蛋白质组。
收集10例牙周炎患者、10例龋齿患者和10例口腔健康个体的刺激唾液样本。唾液样本中的蛋白质用变性缓冲液处理，并用LysC和胰蛋白酶进行酶解。所得肽混合物通过固相萃取进行净化，并通过纳米级C18反相色谱在线分离，梯度为2小时，通过电喷雾源连接到质谱仪。洗脱的肽在以数据依赖采集模式运行的串联质谱仪上进行分析。
我们从4161种不同蛋白质中总共鉴定出35664种独特肽，其中分别有1946种和2090种来自细菌和人类。与健康对照相比，牙周炎和龋齿患者的人类蛋白质谱显示补体系统和炎症标志物明显过表达。健康和疾病状态下的细菌蛋白质组谱和功能注释非常相似。
与补体系统和炎症相关的蛋白质过表达似乎与口腔疾病状态相关。健康和患病个体中相似的细菌蛋白质组表明，唾液微生物群主要以浮游状态生长，不表现出与疾病相关的代谢活性特征。
