Sheep intelectin-2 co-purifies with the mucin Muc5ac from gastric mucus

Secretion of gastric mucins plays an essential role in host protection, and modifications in mucus properties are characteristic of the protective immune responses to pathogens. This study describes the purification and characterisation of sheep gastric mucins, and identification of those proteins that co-purify with mucins, with the potential to modify mucus properties. Gastric mucus was collected and pooled from four abattoir sheep and separated by CsCl density gradient centrifugation. Proteomic analysis of the mucin-containing fraction indicated the presence of gastric mucin (Muc5ac) and several co-purifying proteins, including intelectin-2 (Itln2). Further experimentation indicated that a combination of denaturation and reduction was required to fully release Itln2 from gastric mucin. A putative correlation was found between mucin-bound intelectin concentration and rheological properties in further sheep gastric mucus samples. In conclusion, this study provides the first characterisation of sheep gastric mucins and their purification partners, revealing potentially important mucin-intelectin interactions.