title: Quantitative gingival crevicular fluid proteome in health and periodontal disease using stable isotope chemistries and mass spectrometry.
abstract: OBJECTIVE
  Application of quantitative stable isotope-labelling chemistries and mass spectrometry (MS) to determine alterations in gingival crevicular fluid (GCF) proteome in periodontal disease.
  
  
  METHODS
  Quantitative proteome of GCF from 40 healthy individuals versus 40 patients with periodontal disease was established using 320 GCF samples and stable isotope-labelling reagents, ICAT and mTRAQ, with MS technology and validated by enzyme-linked immunosorbent methods.
  
  
  RESULTS
  We have identified 238 distinct proteins of which 180 were quantified in GCF of both healthy and periodontal patients with additional 26 and 32 distinct proteins that were found only in GCF of healthy or periodontal patients. In addition, 42 pathogenic bacterial proteins and 11 yeast proteins were quantified. The data highlighted a series of proteins not quantified previously by large-scale MS approaches in GCF with relevance to periodontal disease, such as host-derived Ig alpha-2 chain C, Kallikrein-4, S100-A9, transmembrane proteinase 13, peptidase S1 domain, several collagen types and pathogenic bacterial proteins, e.g. formamidase, leucine aminopeptidase and virulence factor OMP85.
  
  
  CONCLUSIONS
  The innovative analytical approaches provided detailed novel changes in both host and microbial derived GCF proteomes of periodontal patients. The study defined 50 host and 16 pathogenic bacterial proteins significantly elevated in periodontal disease most of which were novel with significant potential for application in the clinical arena of periodontal disease.
authors.affiliation: Department of Periodontology and Oral Biology, School of Dental Medicine, Boston University, Boston, MA, USA.
authors.forename: Leandro G
authors.initials: LG
authors.lastname: Carneiro
authors.forename: Hesham
authors.initials: H
authors.lastname: Nouh
authors.forename: Erdjan
authors.initials: E
authors.lastname: Salih
mesh_terms: D000328:Adult; D000418:Albumins / Q000032:analysis; D000581:Amidohydrolases / Q000032:analysis; D001425:Bacterial Outer Membrane Proteins / Q000032:analysis; D001426:Bacterial Proteins / Q000032:analysis; D040502:Calgranulin B / Q000032:analysis; D002853:Chromatography, Liquid; D003094:Collagen / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005656:Fungal Proteins / Q000032:analysis; D005883:Gingival Crevicular Fluid / Q000737:chemistry*; D006801:Humans; D007137:Immunoglobulin alpha-Chains / Q000032:analysis; D007554:Isotopes; D007610:Kallikreins / Q000032:analysis; D007931:Leucyl Aminopeptidase / Q000032:analysis; D008297:Male; D013058:Mass Spectrometry; D008565:Membrane Proteins / Q000032:analysis; D008875:Middle Aged; D010510:Periodontal Diseases / Q000378:metabolism* / Q000382:microbiology; D020543:Proteome / Q000032:analysis*; D012697:Serine Endopeptidases / Q000032:analysis; D012709:Serum Albumin / Q000032:analysis; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry; D055815:Young Adult
chemical_list: D000418:Albumins; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D040502:Calgranulin B; D005656:Fungal Proteins; D007137:Immunoglobulin alpha-Chains; D007554:Isotopes; D008565:Membrane Proteins; D020543:Proteome; D012709:Serum Albumin; D003094:Collagen; D007931:Leucyl Aminopeptidase; D007610:Kallikreins; D012697:Serine Endopeptidases; C429035:TMPRSS13 protein, human; C120801:kallikrein 4; D000581:Amidohydrolases; C101846:formamidase
keywords: biomarkers; diagnostics; gingival crevicular fluid; mass spectrometry; periodontal disease; quantitative proteomics; stable isotope labelling
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