Top-down HPLC-ESI-MS proteomic analysis of saliva of edentulous subjects evidenced high levels of cystatin A, cystatin B and SPRR3.
OBJECTIVE
This study aims to analyze the salivary peptidome/proteome of edentulous subject with respect to dentate control subjects.


METHODS
Unstimulated whole saliva, collected from 11 edentulous subjects (age 60-76 years) and 11 dentate age-matched control subjects, was immediately treated with 0.2% aqueous trifluoroacetic acid and the acidic soluble fraction analyzed by High Performace Liquid Chromatography-Mass Spectrometry. The relative abundance of the salivary peptides/proteins was determined by measuring the area of the High Performace Liquid Chromatography-Mass Spectrometry eXtracted Ion Current peaks which is linearly proportional to peptide/protein concentration under identical experimental conditions. Levels of salivary peptides/proteins in the two groups were compared by the nonparametric Mann-Whitney test to evidence statistically significant differences.


RESULTS
Levels of cystatin A, S-glutathionylated, S-cystenylated, S-S dimer derivatives of cystatin B and S-glutathionylated derivative of SPRR3, were found significantly higher in edentulous subjects with respect to dentate controls. The major peptides and proteins typically deriving from salivary glands did not show any statistically significant differences.


CONCLUSIONS
Cystatin A, S-glutathionylated, S-cystenylated, S-S dimer derivatives of cystatin B and S-glutathionylated derivative of SPRR3, which are mainly of intracellular origin and represent the major constituents of the cornified cell envelope are a clue of inflammation of mucosal epithelia.
Dipartimento di Scienze della Vita e dell'Ambiente, Università di Cagliari, 09042, Monserrato CA, Italy. Electronic address: bmanconi@unica.it.
Barbara
B
Manconi
Dipartimento di Scienze della Vita e dell'Ambiente, Università di Cagliari, 09042, Monserrato CA, Italy.
Barbara
B
Liori
Dipartimento di Scienze della Vita e dell'Ambiente, Università di Cagliari, 09042, Monserrato CA, Italy.
Tiziana
T
Cabras
Istituto di Biochimica e Biochimica Clinica, Università Cattolica, L.go F. Vito 1, 00168 Roma, Italy.
Federica
F
Iavarone
Istituto di Clinica Odontoiatrica, Università Cattolica, L.go F. Vito 1 00168, Roma, Italy.
Armando
A
Manni
Istituto di Chimica del Riconoscimento Molecolare - UOS Roma, Consiglio Nazionale delle Ricerche, Rome, Italy; Facoltà di Scienze Mediche, LUdeS Foundation H.E.I., Lugano, Switzerland.
Irene
I
Messana
Istituto di Biochimica e Biochimica Clinica, Università Cattolica, L.go F. Vito 1, 00168 Roma, Italy; Istituto di Chimica del Riconoscimento Molecolare - UOS Roma, Consiglio Nazionale delle Ricerche, Rome, Italy.
Massimo
M
Castagnola
Dipartimento di Scienze della Vita e dell'Ambiente, Università di Cagliari, 09042, Monserrato CA, Italy.
Alessandra
A
Olianas
D000368:Aged; D016022:Case-Control Studies; D002851:Chromatography, High Pressure Liquid; D055233:Cornified Envelope Proline-Rich Proteins / Q000378:metabolism*; D055312:Cystatin A / Q000378:metabolism*; D055313:Cystatin B / Q000378:metabolism*; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D009066:Mouth, Edentulous; D040901:Proteomics / Q000379:methods*; D012463:Saliva / Q000737:chemistry*; D021241:Spectrometry, Mass, Electrospray Ionization
D055233:Cornified Envelope Proline-Rich Proteins; D055312:Cystatin A; C112254:SPRR3 protein, human; D055313:Cystatin B
Cystatins; Edentulous subjects; Keratinocytes; Mass-Spectrometry; Proteomics; Saliva