Characterization of two isoforms of human SPRR3 from saliva of preterm human newborn and autoptic fetal oral mucosa, parotid and submandibular gland samples

MANCONI, BARBARA;CABRAS, TIZIANA;Pisano E;Nemolato S;Inzitari R;Iavarone F;Fanali C;SANNA, MARIA TERESA;Tirone C;Vento G;Romagnoli C;FAA, GAVINO;Castagnola M;MESSANA, IRENE

2010-01-01

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Abstract

RP-HPLC-ESI-MS profile of saliva samples from human preterm newborn showed a protein peak in the elution range 26.6-27.6 min. Deconvolution of ESI-MS spectra revealed the presence of two proteins with average molecular mass (May) values of 17,239 +/- 3 Da and 18,065 +/- 3 Da in 9 samples, with May value of 17,239 +/- 3 Da in 4 samples and May value of 18,065 +/- 3 Da in 2 samples. MALDI-TOF-MS analysis of tryptic digest allowed identifying the proteins as two isoforms of small proline-rich protein 3 and cDNA amplification of RNA extracts from oral mucosa, parotid and submandibular gland samples, obtained at fetal autopsy, provided two nucleotide sequences in agreement with those reported in the literature. The two proteins differ for an octapeptide repeat (GCTKVPEP) and the substitution Leu -> Val, at position 148 and 140 of the mature form of the 18,065 and 17,239 Da protein, respectively. During maturation the two proteins undergo two post-translational modifications, corresponding to N-terminal acetylation and removal of the initiator methionine. cDNA amplification did not allow to clarify if the proteins found in saliva originated from cellular shedding of the epithelium and/or secretion.