Yuca Esra, Şeker Urartu Özgür Şafak
Molecular Biology and Genetics Department, Yildiz Technical University, İstanbul, Turkey.
UNAM-Institute of Materials Science and Nanotechnology, Bilkent University, Ankara, Turkey.
Methods Mol Biol. 2022;2538:25-33. doi: 10.1007/978-1-0716-2529-3_3.
The structure and the functionality of biofilm proteins, the main components of the extracellular matrix, can be tuned by protein engineering. The use of binding kinetics data has been demonstrated in the characterization of recombinantly produced biofilm proteins to control their behavior on certain surfaces or under certain conditions. Quartz crystal microbalance with dissipation monitoring (QCM-D) allows measuring the change in resonance frequency and the energy loss and distribution upon the interaction of molecules with the surface. The characterization of the molecular assembly of curli biofilm proteins on different surfaces using QCM-D is presented here as a detailed protocol. The experimental procedure detailed in this chapter can be applied and modified for other biofilm proteins or subunits to determine their surface adsorption and kinetic binding characteristics.
生物膜蛋白作为细胞外基质的主要成分,其结构和功能可通过蛋白质工程进行调控。结合动力学数据已被证明可用于表征重组生产的生物膜蛋白,以控制其在特定表面或特定条件下的行为。具有耗散监测功能的石英晶体微天平(QCM-D)能够测量分子与表面相互作用时共振频率的变化以及能量损失和分布情况。本文详细介绍了使用QCM-D表征不同表面上卷曲菌毛生物膜蛋白分子组装的实验方案。本章详述的实验步骤可应用于其他生物膜蛋白或亚基,并进行修改,以确定其表面吸附和动力学结合特性。