Department of Molecular Medicine, University of Padua, Padua, Italy.
Cell Biology and Neuroscience Institute, University of Buenos Aires-National Scientific and Technical Council (UBA-CONICET), Buenos Aires, Argentina.
Sci Rep. 2019 Jan 22;9(1):300. doi: 10.1038/s41598-018-36391-3.
The mouse Major Urinary Proteins (MUPs) contain a conserved β-barrel structure with a characteristic central hydrophobic pocket that binds a variety of volatile compounds. After release of urine, these molecules are slowly emitted in the environment where they play an important role in chemical communication. MUPs are highly polymorphic and conformationally stable. They may be of interest in the construction of biosensor arrays capable of detection of a broad range of analytes. In this work, 14 critical amino acids in the binding pocket involved in ligand interactions were identified in MUP20 using in silico techniques and 7 MUP20 mutants were synthesised and characterised to produce a set of proteins with diverse ligand binding profiles to structurally different ligands. A single amino acid substitution in the binding pocket can dramatically change the MUPs binding affinity and ligand specificity. These results have great potential for the design of new biosensor and gas-sensor recognition elements.
小鼠主要尿蛋白 (MUP) 含有一个保守的β-桶结构，具有特征性的中央疏水性口袋，可结合多种挥发性化合物。尿液排出后，这些分子会在环境中缓慢释放，在其中它们在化学通讯中发挥着重要作用。MUP 具有高度多态性和构象稳定性。它们可能在构建能够检测广泛分析物的生物传感器阵列方面具有重要意义。在这项工作中，使用计算机技术在 MUP20 中鉴定了参与配体相互作用的结合口袋中的 14 个关键氨基酸，并合成了 7 个 MUP20 突变体并进行了表征，以产生一组具有不同配体结合特性的蛋白质与结构不同的配体。结合口袋中的单个氨基酸取代可以极大地改变 MUPs 的结合亲和力和配体特异性。这些结果对于设计新型生物传感器和气体传感器识别元件具有巨大的潜力。
