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唾液处理方法对蛋白质组图谱及蛋白介导的生物学过程的影响

Effect of Processing Methods of Human Saliva on the Proteomic Profile and Protein-Mediated Biological Processes.

机构信息

Department of Prosthodontics and Periodontology, Piracicaba Dental School, University of Campinas (UNICAMP), Piracicaba, Sa̅o Paulo 13414-903, Brazil.

Department of Restorative Dentistry, Federal University of Paraná (UFPR), Curitiba, Paraná 80210-170, Brazil.

出版信息

J Proteome Res. 2023 Mar 3;22(3):857-870. doi: 10.1021/acs.jproteome.2c00652. Epub 2023 Feb 13.

DOI:10.1021/acs.jproteome.2c00652
PMID:36779809
Abstract

The use of saliva as a protein source prior to microbiological and biological assays requires previous processing. However, the effect of these processing methods on the proteomic profile of saliva has not been tested. Stimulated human saliva was collected from eight healthy volunteers. Non-processed saliva was compared with 0.22 μm filtered, 0.45 μm filtered, and pasteurized saliva, by liquid chromatography-mass spectrometry. Data are available via ProteomeXchange with identifier PXD039248. The effect of processed saliva on microbial adhesion was tested using bacterial and fungus species and in biological cell behavior using HaCaT immortalized human keratinocytes. Two hundred and seventy-eight proteins were identified in non-processed saliva, of which 54 proteins (≈19%) were exclusive. Saliva processing reduced identified proteins to 222 (≈80%) for the 0.22 μm group, 219 (≈79%) for the 0.45 μm group, and 201 (≈72%) for the pasteurized saliva, compared to non-processed saliva. The proteomic profile showed similar molecular functions and biological processes. The different saliva processing methods did not alter microbial adhesion (ANOVA, > 0.05). Interestingly, pasteurized saliva reduced keratinocyte cell viability. Saliva processing methods tested reduced the proteomic profile diversity of saliva but maintained similar molecular functions and biological processes, not interfering with microbial adhesion and cell viability, except for pasteurization, which reduced cell viability.

摘要

在进行微生物学和生物学检测之前,使用唾液作为蛋白质来源需要进行预处理。然而,这些处理方法对唾液蛋白质组图谱的影响尚未经过测试。从 8 名健康志愿者中采集了未处理的人唾液。通过液相色谱-质谱法,将非处理的唾液与 0.22μm 过滤、0.45μm 过滤和巴氏消毒的唾液进行了比较。数据可通过 ProteomeXchange 以标识符 PXD039248 获得。使用细菌和真菌物种测试了处理后的唾液对微生物黏附的影响,并使用 HaCaT 永生化人角质形成细胞测试了其对生物细胞行为的影响。在未处理的唾液中鉴定出 278 种蛋白质,其中 54 种(≈19%)是特有的。与非处理的唾液相比,0.22μm 组的鉴定蛋白减少到 222(≈80%),0.45μm 组减少到 219(≈79%),巴氏消毒组减少到 201(≈72%)。蛋白质组图谱显示了相似的分子功能和生物学过程。不同的唾液处理方法并未改变微生物黏附(ANOVA,>0.05)。有趣的是,巴氏消毒降低了角质形成细胞的活力。测试的唾液处理方法降低了唾液的蛋白质组图谱多样性,但保持了相似的分子功能和生物学过程,除了巴氏消毒会降低细胞活力外,不会干扰微生物黏附和细胞活力。

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