Different isoforms and post-translational modifications of human salivary acidic proline-rich proteins

INZITARI R;CABRAS, TIZIANA;ONNIS G;OLMI C;MASTINU, ANDREA;SANNA, MARIA TERESA;PELLEGRINI MG;CASTAGNOLA M;MESSANA, IRENE

2005-01-01

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Abstract

The human salivary acidic proline-rich proteins (aPRPs) complex was investigated by different chromatographic and mass spectrometric approaches and the main aPRPs, namely PRP-1, PRP2 and PIF-s (15 515 amu), Db-s (17 632 amu) and Pa (15 462 amu) proteins, were detected. All these isoforms are phosphorylated at Ser-8 and Ser-22 and have a pyroglutamic moiety at the N-terminus. Apart from Pa, all the other aPRPs undergo a proteolytic cleavage at Arg-106 residue (Arg-127 in Db-s protein), that generates the small PC peptide (4371 amu) and PRP-3, PRP-4, PIF-f (11 162 amu) and Db-f (13 280 amu) proteins, all of which were detected. With regard to the Pa protein, the main form detected was the dimeric derivative (Pa 2-mer, 30 922 amu) originated by a disulfide bond involving Cys-103 residue. Besides these known isoforms, several previously undetected aPRP derivatives were found (in minor amounts): (i) the triphosphorylated derivatives of PRP-1/PRP-2/PIF-s and Db-s, showing the additional phosphate group at Ser-17; (ii) the mono-phosphorylated forms at either Ser-22 or Ser-8 of PRP-1/PRP-2/PIF-s, PRP-3/PRP-4/P1F-f Db-s and Db-f, (iii) a nonphosphorylated form of PRP-3/PRP-4/PlF-f, (iv) the triphosphorylated and diphosphorylated forms of Pa 2-mer. Moreover, minor quantities of PRP-3/PRP-4/PIF-f lacking the C-terminal Arg (11 006 amu), and of Pa 2-mer lacking the C-terminal Gln (30 793 amu) were found. By this approach the different phenotypes of PRH1 locus in 59 different subjects were characterized.