Analysis of post-translational modifications in soluble proteins involved in chemical communication from mammals and insects.

Different soluble, small polypeptide molecules isolated from biological tissues/body fluids of mammals and insects have been reported to bind odorant compounds and pheromones, and have been accordingly named odorant-binding proteins, pheromone-binding proteins and chemosensory proteins. Mammal and insect odorant-binding and pheromone-binding proteins show evident differences in their primary structure; in insects, the same condition occurs for odorant- and pheromone-binding proteins, which belong to the same protein family, and chemosensory proteins. In order to understand their molecular characteristics, these proteins have been subjected to an extensive analysis for their post-translational modifications through mass spectrometry approaches. Depending on proteins, they have been characterized as having specific disulfide bridge arrangements, which were consistent among molecular sequence homologs. Only selected odorant-binding proteins from mammals have been demonstrated bearing N-linked glycan structures and other modifications. In this review article, we discuss the application of mass spectrometry procedures for the analysis of post-translational modifications in odorant-binding proteins, pheromone-binding proteins and chemosensory proteins. Different example proteins are reported, and protocols are presented for obtaining definitive information in this context.