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Annotation of the human odontoblast cell layer and dental pulp proteomes and N-terminomes Abbey, Simon

Abstract

The proteome and N-terminome of the human odontoblast cell layer was identified for the first time by shotgun proteomic and terminal amine isotopic labeling of substrates (TAILS) N terminomic analyses, respectively, and compared with that of human dental pulp stroma from 3rd molar teeth. After reverse-phase liquid chromatography-tandem mass spectrometry, >170,000 spectra from the shotgun and TAILS analyses were matched by four search engines to 4,888 and 12,063 peptides in the odontoblast cell layer and pulp stroma, respectively. Using the Trans-Proteomic Pipeline, I identified 895 and 2,423 unique proteins in these tissues at an FDR of ≤ 1 %. In the odontoblast cell layer proteome I found proteomic evidence for dentin sialophosphoprotein, which is cleaved into dentin phosphoprotein and dentin sialoprotein, proteins that are important in dentin mineralization. Further, 222 proteins of the odontoblast cell layer were not found in the pulp, suggesting many of these proteins are synthesized preferentially by odontoblasts. I also found minor differences in the odontoblast cell layer between the dental pulp proteomes of older and younger donors. The human dental pulp stroma proteome was expanded by 974 new proteins, not previously identified among the 4,123 proteins identified in our previous dental pulp study (Eckhard et al., 2015). Thus, by exploring the proteome of the odontoblast cell layer and expanding the known dental pulp proteome, we found distinct proteome differences when compared with each other and with dentin. The mass spectrometry raw data and metadata have been deposited to ProteomeXchange with the PXD identifier ˂PXD006557˃.

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Attribution-NonCommercial-NoDerivatives 4.0 International