The Human Odontoblast Cell Layer and Dental Pulp Proteomes and N-Terminomes

The Human Odontoblast Cell Layer and Dental Pulp Proteomes and N-Terminomes

The proteome and N-terminome of the human odontoblast cell layer were 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 26 third molar teeth. After...

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Bibliographic Details
Published in:Journal of dental research Vol. 97; no. 3; pp. 338 - 346
Main Authors: Abbey, S.R., Eckhard, U., Solis, N., Marino, G., Matthew, I., Overall, C.M.
Format: Journal Article
Language:English
Published: Los Angeles, CA SAGE Publications 01-03-2018
SAGE PUBLICATIONS, INC
Subjects:
Biology
Dental pulp
Dentin
Dentin sialoprotein
Dentistry
Dspp protein
Liquid chromatography
Mass spectroscopy
Medical research
N-Terminus
Odontoblasts
Proteins
Proteolysis
Proteomes
Proteomics
Radioactive labeling
Stroma
Tails
Teeth
dentin sialophosphoprotein
dentin
proteomics
TAILS
extracellular matrix
N-terminomics
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Summary:The proteome and N-terminome of the human odontoblast cell layer were 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 26 third molar teeth. After reverse-phase liquid chromatography–tandem mass spectrometry, >170,000 spectra from the shotgun and TAILS analyses were matched by 4 search engines to 4,888 and 12,063 peptides in the odontoblast cell layer and pulp stroma, respectively. Within these peptide groups, 1,543 and 5,841 protein N-termini, as well as 895 and 2,423 unique proteins, were identified with a false discovery rate of ≤1%. Thus, the human dental pulp proteome was expanded by 974 proteins not previously identified among the 4,123 proteins in our 2015 dental pulp study. Further, 222 proteins of the odontoblast cell layer were not found in the pulp stroma, suggesting many of these proteins are synthesized only by odontoblasts. When comparing the proteomes of older and younger donors, differences were more apparent in the odontoblast cell layer than in the dental pulp stroma. In the odontoblast cell layer proteome, we found proteomic evidence for dentin sialophosphoprotein, which is cleaved into dentin sialoprotein and dentin phosphoprotein. By exploring the proteome of the odontoblast cell layer and expanding the known dental pulp proteome, we found distinct proteome differences compared with each other and with dentin. Moreover, between 61% and 66% of proteins also occurred as proteoforms commencing with a neo–N-terminus not annotated in UniProt. Hence, TAILS increased proteome coverage and revealed considerable proteolytic processing, by identifying stable proteoforms in these dynamic dental tissues. All mass spectrometry raw data have been deposited to ProteomeXchange with the identifier , with the accompanying metadata at Mendeley Data (https://data.mendeley.com/datasets/b57zfh6wmy/1).
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ISSN:0022-0345
1544-0591
DOI:10.1177/0022034517736054