Identification of protein components in human acquired enamel pellicle and whole saliva using novel proteomics approaches.
Precursor proteins of the acquired enamel pellicle derive from glandular and non-glandular secretions, which are components of whole saliva. The purpose of this investigation was to gain further insights into the characteristics of proteins in whole saliva and in vivo formed pellicle components. To maximize separation and resolution using only micro-amounts of protein, a two-dimensional gel electrophoresis system was employed. Protein samples from parotid secretion, submandibular/sublingual secretion, whole saliva, and pellicle were subjected to isoelectric focusing followed by SDS-PAGE. Selected protein spots were excised, subjected to "in-gel" trypsin digestion, and examined by mass spectrometry (MS). The data generated, including peptide maps and tandem MS spectra, were analyzed using protein data base searches. Components identified in whole saliva include cystatins (SA-III, SA, and SN), statherin, albumin, amylase, and calgranulin A. Components identified in pellicle included histatins, lysozyme, statherin, cytokeratins, and calgranulin B. The results showed that whole saliva and pellicle have more complex protein patterns than those of glandular secretions. There are some similarities and also distinct differences between the patterns of proteins present in whole saliva and pellicle. MS approaches allowed identification of not only well characterized salivary proteins but also novel proteins not previously identified in pellicle.
Yao
Yuan
Y
Department of Periodontology and Oral Biology, Boston University School of Dental Medicine, Boston University School of Medicine, Boston, Massachusetts 02118, USA.
Berg
Eric A
EA
Costello
Catherine E
CE
Troxler
Robert F
RF
Oppenheim
Frank G
FG
D003746:Dental Enamel Proteins / Q000032:analysis*; D044622:Dental Pellicle; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D011498:Protein Precursors / Q000032:analysis; D040901:Proteomics / Q000379:methods; D012471:Salivary Proteins and Peptides / Q000032:analysis*; D012680:Sensitivity and Specificity
D003746:Dental Enamel Proteins; D011498:Protein Precursors; D012471:Salivary Proteins and Peptides