Trial Proteomic Qualitative and Quantitative Analysis of the Protein Matrix of Submandibular Sialoliths

Abstract

Our studies aimed to explore the protein components of the matrix of human submandibular gland sialoliths. A qualitative analysis was carried out based on the filter aided sample preparation (FASP) methodology. In the protein extraction process, we evaluated the applicability of the standard demineralization step and the use of a lysis buffer containing sodium dodecyl sulfate (SDS) and dithiothreitol (DTT). The analysis of fragmentation spectra based on the human database allowed for the identification of 254 human proteins present in the deposits. In addition, the use of multi-round search in the PEAKS Studio program against the bacterial base allowed for the identification of 393 proteins of bacterial origin present in the extract obtained from sialolith, which so far has not been carried out for this biological material. Furthermore, we successfully applied the SWATH methodology, allowing for a relative quantitative analysis of human proteins present in deposits. The obtained results correlate with the classification of sialoliths proposed by Tretiakow. The performed functional analysis allowed for the first time the selection of proteins, the levels of which differ between the tested samples, which may suggest the role of these proteins in the calcification process in different types of sialoliths. These are preliminary studies, and drawing specific conclusions requires research on a larger group, but it provides us the basis for the continuation of the work that has already begun.

Proteomic analysis of sialoliths from calcified, lipid and mixed groups as a source of potential biomarkers of deposit formation in the salivary glands

“…The used approaches treating salivary stone samples were based on protocols described in the publication with the results of trial proteomic analysis of sialoliths [ 48 ]. The first step included crushing the stones into the powder in a mortar.…”

“…Our first published research [ 48 ], based on a relatively small sialoliths group (five stones), showed some proteins with a critical role during the formation of sialoliths. One of them was a group of cystatins (CYST1-5).…”

Proteomic analysis of sialoliths from calcified, lipid and mixed groups as a source of potential biomarkers of deposit formation in the salivary glands

“…The used approaches treating salivary stone samples were based on protocols described in the publication with the results of trial proteomic analysis of sialoliths [51]. The rst step included crushing the stones into the powder in a mortar.…”

“…There were detected precisely the same species: Actinomyces viscosus, Eikenella corrodens, Fusobacterium nucleatum, Haemophilus parain uenzae and Streptococcus mitis. What is more, identi ed bacteria, if they were not the same species, they were from the same groups: Actinomyces, Capnocytophaga, Eikenella, Haemophylus, Kingella, Neisseria, Peptostreptococcus, Porphyromonas, Rothia and Streptococcus [22], [45], [49]- [51]. All of the identi ed bacteria groups are common in the oral cavity.…”

“…Our rst published research [51], based on a relatively small sialoliths group ( ve stones), showed some proteins with a critical role during the formation of sialoliths. One of them was a group of cystatins (CYST1-5).…”