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Proteomic study of the dentin-enamel junction layer.
Kolrosová, Marta ; Jágr, Michal (advisor) ; Ergang, Peter (referee)
This thesis deals with the analysis of the dentin-enamel junction layer in human third molar teeth. Three different methods of analysis were tested. In the first one teeth were demineralized in EDTA buffer to release the protein layer DEJ. In the other two cases thin slils of dental tissue were preparedand from them the DEJ layer was cut using laser microdissection. Severals methods were tested for extraction of proteins from DEJ samples (for comparsio also from dentin and enamel samples). Extracted proteins were cleavaged into peptides with trypsin. Subsequently, peptides were purified by Stage tips and analyzed by nLC MS/ MS. Using the optimal method over 40 different proteins, for example: apolipoproteins, vimentin, vitronectin, clusterin, biglycan weres found in the sample DEJ. Keywords: proteomics, dentin-enamel junction, laser microdissection, mass spectrometry
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Proteomic study of human tooth dentin
Zettlová, Ludmila ; Jágr, Michal (advisor) ; Vrbacký, Marek (referee)
Proteome analysis of dentin in human teeth adult subjects was conducted to optimize the two dimensional gel electrophoresis (2-DE) analysis of proteins from the dental tissue. The third molar teeth were cleaned, crushed and extracted in guanidine and EDTA buffer. Extracted proteins were separated by 2-DE, digested with trypsin to peptides and subsequently analyzed using nLC MS/MS. Totally, 7 unique proteins were identified in the samples. The use of IPG strips with different range of the pH gradient and the subsequent comparison of 2 DE gels unfortunately did not bring important new information, because common keratin, collagen and serum albumin proteins were identified.
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