Comparison of protein profiles of the pellicle, gingival crevicular fluid, and saliva: possible origin of pellicle proteins
Comparison of protein profiles of the pellicle, gingival crevicular fluid, and saliva: possible origin of pellicle proteins
复制标题
DOI：
10.1186/s40659-020-0271-2
复制
发表时间：
2020-01-01
影响因子：
6.7
通讯作者：
Yamamoto, Matsuo
中科院分区：
文献类型：
--
作者：
Odanaka, Hibiki;Obama, Takashi;Yamamoto, Matsuo
研究背景牙釉质表面沉积的无细胞有机物质--牙膜被认为来源于唾液。在这项研究中，比较了从健康成人采集的牙周膜、牙周沟液和唾液的蛋白质组成，以阐明膜蛋白的来源。结果分别从腮腺和混合腺采集唾液、牙周膜、龈沟液，用SDS-PAGE和MS分析比较不同个体样本的蛋白质表达。经SDS-PAGE后，用质谱仪鉴定主要条带中的蛋白质。膜蛋白的带型与牙周沟液或唾液的带型不同。利用质谱仪对样品中的13种蛋白质进行了鉴定。结合稳定同位素标记和蛋白质印迹对蛋白质的相对丰度进行了定量分析。蛋白质印迹法检测到膜中的半胱氨酸蛋白酶、S和α-淀粉酶在唾液中含量较高，而在牙周沟液中未检测到；稳定同位素标记分析表明，它们在唾液中的丰度较高，在牙周沟液中的丰度较低。免疫印迹法检测血清转铁蛋白仅见于牙周膜和牙周沟液中，唾液中血清转铁蛋白丰度较低。结论我们的研究表明，除唾液外，牙周沟液似乎也对透明膜的形成有贡献。
Background The pellicle, the acellular organic material deposited on the surface of tooth enamel, has been thought to be derived from saliva. In this study, protein compositions of the pellicle, gingival crevicular fluid, and saliva collected from healthy adults were compared to elucidate the origin of pellicle proteins. Results The pellicle, gingival crevicular fluid, and saliva from the parotid gland or mixed gland were collected; subsequently, protein expression in samples from the respective individual was compared by SDS-PAGE and mass spectrometry. Following SDS-PAGE, proteins in the major bands were identified by mass spectrometry. The band pattern of pellicle proteins appeared different from those of gingival crevicular fluid, or saliva samples. Using mass spectrometry, 13 proteins in these samples were identified. The relative abundance of the proteins was quantitatively analyzed using mass spectrometry coupled with stable isotope labeling and by western blot. Cystatin S and alpha-amylase detected in pellicle were enriched in saliva samples, but not in gingival crevicular fluid, by western blot, and their abundance ratios were high in saliva and low in gingival crevicular fluid when analyzed by stable isotope labeling. Serotransferrin, however, was found only in the pellicle and gingival crevicular fluid by western blot and its abundance ratio was low in saliva. Conclusions Our study revealed that the gingival crevicular fluid appears to contribute to pellicle formation in addition to saliva.