| Title: | Molecular and functional characterization of buffalo nasal epithelial odorant binding proteins and their structural insights by in silico and biochemical approaches |
| Author(s): | Manikkaraja C; Bhavika M; Singh R; Nagarathnam B; George G; Gulyani A; Archunan G; Sowdhamini R; |
| Address: | "Pheromone Technology Lab, Department of Animal Science, Bharathidasan University, Tiruchirappalli, Tamil Nadu, India. GKVK Campus, National Centre for Biological Sciences, Tata Institute for Fundamental Research, Bangalore, Karnataka, India. The University of Trans-Disciplinary Health Sciences and Technology (TDU), Bangalore, Karnataka, India. The Institute for Stem Cell Biology and Regenerative Medicine, National Centre for Biological Sciences (NCBS), Bangalore, Karnataka, India. Department of Biochemistry, School of Life Sciences, University of Hyderabad, Hyderabad, Telangana, India" |
| DOI: | 10.1080/07391102.2020.1854117 |
| ISSN/ISBN: | 1538-0254 (Electronic) 0739-1102 (Linking) |
| Abstract: | "The olfactory system is capable of detecting and distinguishing thousands of environmental odorants that play a key role in reproduction, social behaviours including pheromones influenced classical events. Membrane secretary odorant binding proteins (OBPs) are soluble lipocalins, localized in the nasal membrane of mammals. They bind and carry odorants within the nasal epithelium to putative olfactory transmembrane receptors (ORs). OBP has not yet been exploited to develop a suitable technique to detect oestrus which is being reported as a difficult task in buffalo. In the present study, using molecular biology and protein engineering approaches, we have cloned six novel OBP isoforms from buffalo nasal epithelium odorant-binding proteins (bnOBPs). Furthermore, 3 D models were developed and molecular-docking, dynamics experiments were performed by in silico approaches. In particular, we found four residues (Phe104, Phe134, Phe69 and Asn118) in OBP1a, which contributed to favourable interactions towards two sex pheromones, specifically oleic acid and p-cresol. We expressed this protein in Escherichia coli from female buffalo urine and validated through fluorescence quenching studies to show similar strong binding affinities of OBP1a to oleic acid and p-cresol. By using structural data, the binding specificity was also verified by site-directed mutagenesis of the four residues followed by in vitro binding assays. Our results enable us to better understand the functions of different nasal epithelium OBP isoforms in buffaloes. They also lead to improved understanding of the interaction between olfactory proteins and odorants to develop highly selective biosensing devices for non-invasive detection of oestrus in buffaloes. Communicated by Ramaswamy H. Sarma" |
| Keywords: | "Animals *Buffaloes/metabolism Female Molecular Docking Simulation Odorants Oleic Acid Protein Isoforms *Receptors, Odorant/chemistry/genetics/metabolism Buffalo chemical communications competitive binding assay molecular docking odorant binding proteins s;" |
| Notes: | "MedlineManikkaraja, Chidhambaram Bhavika, Mam Singh, Randhir Nagarathnam, Balasubramanian George, Geen Gulyani, Akash Archunan, Govindaraju Sowdhamini, Ramanathan eng Research Support, Non-U.S. Gov't England 2020/12/10 J Biomol Struct Dyn. 2022 Jun; 40(9):4164-4187. doi: 10.1080/07391102.2020.1854117. Epub 2020 Dec 9" |
