Dataset Information

Effect of processing methods of human saliva on proteomic profile and protein-mediated biological processes

ABSTRACT: Human saliva has been commonly used as protein source in in vitro microbiological and biological assays to mimic the protein pellicle formation, termed acquired salivary pellicle, that precedes microbial and cell adhesion on surfaces exposed to the oral environment. However, saliva requires previous processing to remove food debris, microorganisms, and other molecules prior its use in microbiological and biological in vitro assays. For this purpose, 0.22 μm filtration, 0.45 μm filtration, and pasteurization methods have been commonly used, but the effect of these processing methods on the proteomic profile of saliva has not been tested experimentally. Stimulated human saliva was collected from 8 healthy volunteers and submitted to the following processes: non-processing (control), 0.22 μm filtration, 0.45 μm filtration, and pasteurization. The proteomic profile of non-processed saliva was compared with 0.22 μm filtered-, 0.45 μm filtered-, and pasteurized-saliva by liquid chromatography-mass spectrometry. The effect of processed saliva in microbial adhesion was tested using bacterial and fungus species, and in biological cell behavior using HaCaT immortalized human keratinocytes. Two hundred seventy-eight proteins were identified in non-processed saliva, 54 proteins (≈19%) were exclusive. Saliva processing reduced identified proteins to 222 (≈80%) for the 0.22 μm filtered saliva, 219 (≈79%) for the 0.45 μm filtered saliva, and 201 (≈72%) for the pasteurized saliva, compared to non-processed saliva. Although there were slight differences in the protein composition, the proteomic profile showed similar molecular functions and biological processes. The different saliva processing methods did not alter microbial adhesion (ANOVA, p>0.05). Interestingly, pasteurized saliva reduced keratinocytes cell viability. Saliva processing methods tested reduced the proteomic profile diversity of saliva, but maintained similar molecular functions and biological processes mediated by remaining proteins, not interfering on microbial adhesion and cell viability, except for pasteurization, which reduced cell viability.

INSTRUMENT(S): LTQ Orbitrap

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Saliva

SUBMITTER: Luis Fernando Bandeira Miranda

LAB HEAD: João Gabriel S. Souza

PROVIDER: PXD039248 | Pride | 2023-03-11

REPOSITORIES: Pride

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Dataset's files

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			Action	DRS
	SEARCH_-_Saliva_Processing_Methods_Proteome.xlsx	Xlsx
	Saliva Processing Methods Proteome - LFQ intensity, Ms Count, and Intensity of Identified Proteins.xml	Xml
	Table_of_Contents.xls	Xls
	checksum.txt	Txt

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Publications

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

Miranda Luis Fernando B LFB Lima Carolina V CV Pagin Rafaela R Costa Raphael C RC Pereira Marta Maria A MMA de Avila Erica D ED Bertolini Martinna M Retamal-Valdes Belén B Shibli Jamil A JA Feres Magda M Barão Valentim A R VAR Souza João Gabriel S JGS

Journal of proteome research 20230213 3

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 ...[more]

PMID: 36779809

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