Identification of mouse submaxillary gland protein in mouse saliva and its binding to mouse oral bacteria

Abstract

The mouse submaxillary gland protein (mSMGP) is highly expressed in the submandibular gland of the adult mouse and rat. It shares 51% identity at the amino-acid level with a human protein, the prolactin-inducible protein (PIP)/gross cystic disease fluid protein 15 (GCDFP-15), which has been found in saliva, tears, sweat, seminal plasma, submucosal glands of the lung and amniotic fluid. More recently, the human PIP has been reported to bind to bacterial strains normally found in the mouth, ear canal and human skin. Sequence analysis of mSMGP/PIP earlier identified the presence of a signal peptide, suggesting that it is a secreted protein. Here, by Western blotting, mSMGP/PIP has been identified in mouse saliva. To investigate further the role of this secreted protein, its ability to bind specifically to oral bacteria was examined; the hypothesis was that mSMGP/PIP is involved in non-immune host defence by binding to bacteria. Several bacterial strains, found to belong to the genera Streptococcus, Aerococcus, Pseudomonas, Staphylococcus, Sphingomonas, Vibrio and Aeromonas, were isolated from the mouse oral cavity. Following incubation of these bacteria with 35S-labeled, in vitro-translated mSMGP/PIP, the protein was found to bind specifically and selectively to several but not all strains tested, showing the highest affinity for the streptococci. The protein also bound specifically to an Aerococcus sp., and a low binding interaction with the Pseudomonas and Staphylococcus spp. was observed. The conservation of SMGP sequences among several animal species suggests that this protein may play an important part in the biology of the submandibular gland. As the function of the mSMGP/PIP is still undetermined, these findings provide insight into a possible involvement of this protein in host defence.