Peptide profile of human acquired enamel pellicle using MALDI tandem MS
- 11 February 2008
- journal article
- research article
- Published by Wiley in Journal of Separation Science
- Vol. 31 (3), 523-537
- https://doi.org/10.1002/jssc.200700486
Abstract
The present study proposes a strategy for human in vivo acquired enamel pellicle (AEP) peptidome characterisation based on sequential extraction with guanidine and TFA followed by MALDI‐TOF/TOF identification. Three different nanoscale analytical approaches were used: samples were subjected to tryptic digestion followed by nano‐HPLC and mass spectrometry (MS and MS/MS) analysis. Undigested samples were analysed by LC‐MS (both linear and reflector modes) and LC‐MS/MS analysis, and samples were subjected to nano‐HPLC followed by on‐plate digestion and mass spectrometry (MS and MS/MS) analysis. The majority of the identifications corresponded to peptide/protein fragments of salivary protein, belonging to the classes: acidic PRPs, basic PRPs, statherin, cystatins S and SN and histatin 1 (all also identified in intact form). Overall, more than 90 peptides/proteins were identified. Results clearly show that peptides with acidic groups are enriched in the TFA fraction while peptides with no acidic or phosphate groups are prevalent on the guanidine extract. Also, phosphorylated peptides were observed mainly on the TFA fraction. Fragments present in the AEP show a predominance of cleavage points located at Arg, Tyr and Lys residues. Obtained data suggest that proteolytic activity could influence AEP formation and composition.Keywords
This publication has 42 references indexed in Scilit:
- Identification of Protein Components in in vivo Human Acquired Enamel Pellicle Using LC−ESI−MS/MSJournal of Proteome Research, 2007
- Detection of salivary α‐amylase and lysozyme exposed on the pellicle formed in situ on different materialsJournal of Biomedical Materials Research Part A, 2007
- Two‐dimensional electrophoresis study of in vitro pellicle formation and dental caries susceptibilityEuropean Journal of Oral Sciences, 2006
- The role of salivary peptides in dental cariesBiomedical Chromatography, 2004
- In vitro hydroxyapatite adsorbed salivary proteinsBiochemical and Biophysical Research Communications, 2004
- Salivary Proteins: Protective and Diagnostic Value in Cariology?Caries Research, 2004
- Cysteine Peptidases of Mammals: Their Biological Roles and Potential Effects in the Oral Cavity and Other Tissues in Health and DiseaseCritical Reviews in Oral Biology & Medicine, 2002
- Electron‐microscopic demonstration of proline‐rich proteins, statherin, and histatins in acquired enamel pelliclesin vitroEuropean Journal of Oral Sciences, 2001
- Rate of pellicle formation in vivoActa Odontologica Scandinavica, 1995
- Basic Biological SciencesJournal of Dental Research, 1980