Site-directed mutagenesis of odorant-binding proteins

Modifying the affinity of odorant-binding proteins (OBPs) to small ligands by replacement of specific residues in the binding pocket may lead to several technological applications. Thanks to their compact and stable structures, OBPs are currently regarded as the best candidates to be used in biosensing elements for odorants and volatiles detection. The wide and rich information on the structure of these proteins both in their apo-forms and in complexes with specific ligands provides guidelines to design reliable mutants to monitor specific targets. The same engineered proteins may also find applications in the slow release of pheromones and other chemicals in the environment, as well as in the fine purification of drugs, including the resolution of racemates. Apart from such useful applications, site-directed mutagenesis represents an interesting approach to dissect the specific interactions between small chemicals and amino acid residues in the binding pocket. These studies can lead to design of better ligands, such as pheromone analogues with desired physico-chemical characteristics. In this chapter we examine the different uses of mutagenesis applied to OBPs and report a couple of protocols that have been successful in our hands.