HPLC-MS characterization of cyclo-statherin Q37, a specific cyclization product of human salivary statherin generated by transglutaminase 2

Emanuele Scarano, Tiziana Cabras, Rosanna Inzitari, Chiara Fanali, Maria Patamia, Maria Teresa Sanna, Elisabetta Pisano, Bruno Giardina, Massimo Castagnola, Irene Messana

Risultato della ricerca: Contributo in rivistaArticolo in rivista

Abstract

In the present study the analytical potential of HPLC-MS/MS was utilized for the structural characterization of a post-translational modification of statherin. Human salivary statherin (M-av 5380.0 +/- 0.3 Da) is transformed by the action of transglutaminase 2 into a cyclic derivative with an average molecular mass of 5363.0 +/- 0.3 Da. The intra-molecular bridge is generated by the loss of an ammonia molecule between the unique lone-pair donating nucleophile Lys-6 and one acceptor among the seven glutamine residues of statherin. Digestion of the cyclic derivative with chymotrypsin, proteinase K, and carboxypeptidase Y, monitored by HPLC - electrospray ionization-ion trap-mass spectrometric analysis, demonstrated that cyclization involved almost specifically Gln-37 (> 95%), with the percentage of Gln-39 implicated in the cross-linking being less than 5%. The main derivative was named cyclo-statherin Q-37. Guinea pig transglutaminase 2 showed high affinity for statherin in vitro (K-m = 0.65 +/- 0.06 mu M). Cyclo-statherin was detected in vivo by HPLC-electros pray ionization ion trap-mass spectrometry analysis of whole human saliva and it accounted for about 1% of total statherin. Detection of cyclo-statherin in whole saliva is suggestive of a putative role of this molecule in the formation of the "oral protein pellicle".
Lingua originaleEnglish
pagine (da-a)2600-2606
Numero di pagine7
RivistaJournal of Separation Science
Stato di pubblicazionePubblicato - 2006

Keywords

  • CYCLIZATION
  • SALIVARY
  • STATHERIN
  • TRANSGLUTAMINASE

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