title: Proteomic analysis of gingival crevicular fluid for discovery of novel periodontal disease markers.
abstract: The protein composition of gingival crevicular fluid (GCF) may reflect the pathophysiology of periodontal diseases. A standard GCF proteomic pattern of healthy individuals would serve as a reference to identify biomarkers of periodontal diseases by proteome analyses. However, protein profiles of GCF obtained from apparently healthy individuals have not been well explored. As a step toward detection of proteomic biomarkers for periodontal diseases, we applied both gel-based and gel-free methods to analyze GCF obtained from healthy subjects as compared with supragingival saliva. To ensure optimized protein extraction from GCF, a novel protocol was developed. The proteins in GCF were extracted with high yield by urea buffer combined with ultrafiltration and the intensity of spots with supragingival saliva and GCF was compared using agarose two-dimensional electrophoresis. Eight protein spots were found to be significantly more intense in GCF. They included superoxide dismutase 1 (SOD1), apolipoprotein A-I (ApoA-I), and dermcidin (DCD). Moreover, GCF proteins from healthy subjects were broken down into small peptide fragments and then analyzed directly by LC-MS/MS analysis. A total of 327 proteins including ApoA-I, SOD1, and DCD were identified in GCF. These results may serve as reference for future proteomic studies searching for GCF biomarkers of periodontal diseases.
authors.affiliation: Department of Molecular Diagnosis, Graduate School of Medicine, Chiba University, Chiba, Japan.
authors.forename: Sachio
authors.initials: S
authors.lastname: Tsuchida
authors.forename: Mamoru
authors.initials: M
authors.lastname: Satoh
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Umemura
authors.forename: Kazuyuki
authors.initials: K
authors.lastname: Sogawa
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Kawashima
authors.forename: Sayaka
authors.initials: S
authors.lastname: Kado
authors.forename: Setsu
authors.initials: S
authors.lastname: Sawai
authors.forename: Motoi
authors.initials: M
authors.lastname: Nishimura
authors.forename: Yoshio
authors.initials: Y
authors.lastname: Kodera
authors.forename: Kazuyuki
authors.initials: K
authors.lastname: Matsushita
authors.forename: Fumio
authors.initials: F
authors.lastname: Nomura
mesh_terms: D000328:Adult; D016632:Apolipoprotein A-I / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism; D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005883:Gingival Crevicular Fluid / Q000737:chemistry*; D006801:Humans; D008297:Male; D010455:Peptides / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism; D010510:Periodontal Diseases / Q000175:diagnosis* / Q000378:metabolism; D011506:Proteins / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D013482:Superoxide Dismutase / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism; D000072105:Superoxide Dismutase-1; D053719:Tandem Mass Spectrometry
chemical_list: D016632:Apolipoprotein A-I; D010455:Peptides; D011506:Proteins; C000606290:SOD1 protein, human; C442243:dermcidin; D013482:Superoxide Dismutase; D000072105:Superoxide Dismutase-1