The high molecular weight mucins with their high degree of O-linked glycosylation (50–80% of total weight) in their Ser/Thr/Pro rich domains [37] is involved in protection against oral bacteria. There is growing evidence that shows that mucin glycosylation can change in response to mucosal infection and inflammation [2]. This

will alter the oral milieu for the bacteria and how they interact Inhibitors,research,lifescience,medical with oral surfaces. Bacteria will degrade oligosaccharides from mucins in order to make them available as a nutrient source [38,39,40]. This degradation is achieved by the production of glycosidases such as; α-N-acetyl-D-galactosamindase, sialidase, β-galactosiminidase, β-N-acetlyglucosaminidase, α-and β-mannosidase, and α-fucosidas [41,42]. The results from salivary MUC5B and MUC7 after incubation with saliva indicate high level of sialidase Inhibitors,research,lifescience,medical activity under the conditions applied. The removal of sialic acid makes new monosaccharide units accessible for salivary exoglycosidases. Hence, this step is important to enable the degradation Inhibitors,research,lifescience,medical of salivary mucins. Preliminary data showed that sialidases and proteases work in parallel to degrade the mucins (data not shown), indicating that sialidase not only exposes new oligosaccharide epitopes for further exoglycosidase digestion, but also makes the protein backbone more accessible for proteolytic degradation. The literature suggests that

the exposure of the mucin protein backbone (mucins expressed in the intestine) to proteolytic enzymes produced by various bacteria [43] may result in the host becoming more prone Inhibitors,research,lifescience,medical to infections, as shown

in the cases of ulcerative colitis and Crohn’s disease [44]. However, the degradation of oral mucins is complex, requiring Sorafenib datasheet multiple strains of bacteria to co-exist Inhibitors,research,lifescience,medical in a symbiotic relationship [45]. Some bacteria produce enzymes that degrade the oligosaccharide side chains of mucins, while others produce proteolytic enzymes [45]. To understand this relationship, measuring the combined effect of multiple exoglycosidases on multiple oligosaccharide epitopes will provide clues into distinguishing the conditions provided by commensal bacteria from pathological conditions. 3. Experimental Section 3.1. Materials and Methods either The sialidase S/NANase I (recombinant from Streptococcus pneumonia, expressed in E. coli), glyko β-N-acetylhexosaminidase (jack bean)/HEXase III, β-N-acetylglucosaminidase (GUH) were obtained from Prozyme Co. (Hayward, CA, USA) and α-N-acetylgalactosaminidase from C. perfringens was obtained from R&D systems (Minneapolis, MN, USA). PGM, dithiothreitol (DTT) and iodoacetamide (IAA) were obtained from Sigma Aldrich Co. (St Louis, MO, USA). Hypersep hypercarb SPE columns (60106-301) were obtained from Thermo Scientific Co. (Sanford, FL, USA). The NuPAGE gels were obtained from Invitrogen Co. (Grand Island, NY, USA). 3.2.