Adsorption of SARS-CoV-2 Spike (N501Y) RBD to Human Angiotensin-Converting Enzyme 2 at a Lipid/Water Interface
Adsorption of SARS-CoV-2 Spike (N501Y) RBD to Human Angiotensin-Converting Enzyme 2 at a Lipid/Water Interface
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DOI：
10.1021/acs.jpcb.3c00832
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发表时间：
2023-05-12
影响因子：
3.3
通讯作者：
Rzeznicka, Izabela I.
中科院分区：
文献类型：
--
作者：
Rozak, Harison;Nihonyanagi, Satoshi;Rzeznicka, Izabela I.
SARS冠状病毒2型（SARS-CoV-2）刺突蛋白的受体结合域（RBD）在SARS冠状病毒2型与人血管紧张素转换酶2（ACE 2）的结合过程中起着重要作用。N501 Y突变和后来的突变在尖峰RBD上引入了额外的正电荷，并导致更高的传递性，这可能是由于与高度负电荷的ACE 2的结合更强。因此，人们对刺突蛋白与ACE 2受体结合的分子机制进行了大量的研究，但大多数理论研究都是在分离蛋白上进行的。ACE 2是一种跨膜蛋白，因此，了解纤突蛋白与ACE 2在脂质基质中的相互作用是非常重要的。本研究采用外差探测振动和频发生（HD-VSFG）技术研究了ACE 2和Spike（N501 Y）RBD在脂/水界面的吸附。该技术是一种非线性光学光谱学，它测量分子在界面处的振动光谱，并提供有关其结构和取向的信息。结果表明，ACE 2通过静电作用被有效吸附在带正电荷的1，2-二棕榈酰-3-三甲基铵-丙烷（DPTAP）脂质单分子膜上。ACE 2在DPTAP单分子膜上的吸附导致界面水（D2 O）从D-down方向重新排列到D-up方向，表明原来带正电荷的DPTAP界面由于ACE 2的吸附而变成带负电荷。带负电荷的界面（DPTAP/ACE 2）允许进一步吸附带正电荷的尖峰RBD。酰胺I区的HD-VSFG光谱显示D2 O、DPTAP和DPTAP/ACE 2界面上吸附的加标（N501 Y）RBD的差异。观察到的光谱的尖峰RBD/DPTAP的红移表明，spikeRBD低聚物与DPTAP脂质接触后形成。
Thereceptor binding domain (RBD) of spike proteins plays a crucialrole in the process of severe acute respiratory syndrome corona virus2 (SARS-CoV-2) attachment to the human angiotensin-converting enzyme2 (ACE2). The N501Y mutation and later mutations introduced extrapositive charges on the spike RBD and resulted in higher transmissibility,likely due to stronger binding with the highly negatively chargedACE2. Consequently, many studies have been devoted to understandingthe molecular mechanism of spike protein binding with the ACE2 receptor.Most of the theoretical studies, however, have been done on isolatedproteins. ACE2 is a transmembrane protein; thus, it is important tounderstand the interaction of spike proteins with ACE2 in a lipidmatrix. In this study, the adsorption of ACE2 and spike (N501Y) RBDat a lipid/water interface was studied using the heterodyne-detectedvibrational sum frequency generation (HD-VSFG) technique. The techniqueis a non-linear optical spectroscopy which measures vibrational spectraof molecules at an interface and provides information on their structureand orientation. It is found that ACE2 is effectively adsorbed atthe positively charged 1,2-dipalmitoyl-3-trimethylammonium-propane(DPTAP) lipid monolayer via electrostatic interactions. The adsorptionof ACE2 at the DPTAP monolayer causes a reorganization of interfacialwater (D2O) from the D-down to the D-up orientation, indicatingthat the originally positively charged DPTAP interface becomes negativelycharged due to ACE2 adsorption. The negatively charged interface (DPTAP/ACE2)allows further adsorption of positively charged spike RBD. HD-VSFGspectra in the amide I region show differences for spike (N501Y) RBDadsorbed at D2O, DPTAP, and DPTAP/ACE2 interfaces. A redshift observed for the spectra of spike RBD/DPTAP suggests that spikeRBD oligomers are formed upon contact with DPTAP lipids.