RP-HPLC-ESI-MS evidenced that salivary cystatin B is detectable in adult human whole saliva mostly as S-modified derivatives: S-Glutathionyl, S-cysteinyl and S-S 2-mer

Federica Iavarone, Emanuele Scarano, Giulio Cesare Passali, Massimo Cordaro, Gaetano Paludetti, Tiziana Cabras, Barbara Manconi, Chiara Fanali, Antonella Fiorita, Armando Manni, Irene Messana, Massimo Castagnola, S Nemolato, G Faa

Risultato della ricerca: Contributo in rivistaArticolo in rivista

26 Citazioni (Scopus)

Abstract

An HPLC-ESI-MS analysis of adult human whole saliva evidenced three protein masses (M average 11,487±2, 11,301±2 and 22,362±3Da) eluting in the 32.5-35.0min range. Treatment in reducing conditions allowed establishing that they are S-derivatives of N-terminal acetylated cystatin B, namely its S-glutathionyl, S-cysteinyl and S-S dimer. The identification was confirmed by high resolution HPLC-ESI-MS-MS experiments on the intact naturally occurring proteins and their tryptic digests. S-unmodified cystatin B is rarely detectable in whole saliva of healthy adults (5 subjects out of 65) and its percentage does not overcome approximately 20% of total cystatin B (11±9%). In the majority of subjects (60 out of 65) the mean percentages of the S-modified derivatives were S-glutathionyl 53±13%, S-cysteinyl 15±5%, S-S 2-mer 32±13%. Variations of the percentages of these S-modified derivatives of cystatin B could be indicative of oral oxidative stress. As we are aware, this is the first time that S-glutathionylation and S-cysteinylation were described as extensive PTM of a salivary protein and the first time that these PTMs were detected in naturally occurring cystatin B.
Lingua originaleEnglish
pagine (da-a)908-913
Numero di pagine6
RivistaJOURNAL OF PROTEOMICS
Volume75
DOI
Stato di pubblicazionePubblicato - 2012

Keywords

  • Adult
  • Chromatography, High Pressure Liquid
  • Cystatin B
  • Female
  • Humans
  • Male
  • Mass Spectrometry
  • Oxidation-Reduction
  • Oxidative Stress
  • Protein Modification, Translational
  • Proteomics
  • Saliva

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