Comprehensive proteomic analysis of human dentin

Abstract

Proteomic analysis of the human body is a significant recent scientific endeavour. In this study, we investigated the proteomic profile of human dentin using modern analytical and mass spectrometric techniques. Five healthy permanent human molars from five adults were cut, pulverized, denaturated with guanidine buffer, and demineralized with EDTA buffer. The extracted proteins were analysed by gel electrophoresis (SDS-PAGE and two-dimensional gel electrophoresis), digested with trypsin, and separated by liquid chromatography/high-resolution tandem mass spectrometry. We identified 289 proteins with high confidence, 90 of which had not been previously detected in human dentin. Nine (currently hypothetical) proteins were identified for the first time in an actual human sample. The proteins have a variety of functions, including calcium-ion binding, formation of the extracellular matrix, formation of the cytoskeleton, cytoskeletal protein binding, immune response, and transport. In conclusion, this is the first use of two-dimensional electrophoresis for investigating human dentin.

Proteomic analysis of human dental cementum and alveolar bone

“…In the present study, denaturant bound-bead-based agitation was used to facilitate release of proteins from ground tissues, without employing sample demineralization. Previous studies analyzing the human dentin proteome have included tissue demineralization, and the resulting total number of proteins detected was found to be 233–269 proteins [22, 23], very similar to the number of proteins detected in the present study, with a total of 235 and 213 proteins for AB and DC samples, respectively.…”

“…In the present study, two selective dentin markers, dentin phosphoprotein (DPP) and dentin sialoprotein (DSP) [22, 23] were not detected indicating that cementum samples were not likely contaminated with dentin. However, periostin (POSTN) was found in AB and DC samples, and its presence might result from traces of PDL contamination to both samples [24].…”

“…These observations may explain a higher expression of one of these proteins to a specific tissue, as in DC compared to AB. Previous studies showed the presence of BGN in dentin [22, 23], mandibular bone cells [22, 36], and dental cementum [37]. The osteoporosis-like phenotype in BGN knock-out mice [36], and its interaction with growth factors including bone morphogenetic protein 4 (BMP-4) [38, 39], underscores the importance of this proteoglycan in the modulation of mineralized tissues.…”

Endocardial Fibroelastosis is Secondary to Hemodynamic Alterations in the Chick Embryonic Model of Hypoplastic Left Heart Syndrome

“…Database searches were performed as described (Jagr et al, 2012), with the taxonomy restricted to Gallus gallus to remove protein identification redundancy. Only significant hits (MASCOT score-80, http://www.matrixscience.com) were accepted.…”

Proteomics and Proteogenomics Approaches for Oral Diseases

“…Few studies, indeed, give detailed pieces of information about oral tissues genomics and proteomics (Jágr et al, 2012). These samples can be collected only in an invasive way, and even though they are quite reliable and reproducible, they are characterized by low acceptance and are therefore challenging.…”