In the realm of insect olfaction, Odorant Binding Proteins (OBPs) play a pivotal role in the capture and transfer of hydrophobic volatile compounds from the ambient air to receptors inside the sensory organs. To study these proteins, they are often expressed in bacterial hosts, which is then almost invariably followed by in-vitro refolding to give the proteins their proper structural conformation and therefore functionality. However, despite their critical role, a limited number of comprehensive studies have systematically explored and compared different refolding methodologies for the insect OBPs, often leaving questions regarding the effectiveness of these approaches unanswered.
This research seeks to address this knowledge gap by conducting a thorough comparative analysis of various refolding methods, including four well-established techniques and a novel approach developed in this study. The study centres on three distinct OBPs derived from Helicoverpa armigera, namely HarmOBP2, HarmOBP5, and HarmGOBP2, selected as representative models. To our knowledge, this is the first comprehensive examination of insect OBP refolding, coupled with structural characterization. The results underscore the significance of understanding and optimizing the refolding of recombinant OBP, thereby enhancing the reliability and applicability of these proteins in diverse scientific and technological contexts.
In conclusion, this research contributes to our understanding of insect OBP refolding, emphasizing the importance of methodological scrutiny, and promises to facilitate the utilization of these proteins in future applications while shedding light on the fascinating mechanisms underlying insect sensory perception.