Alimova Alexandra, Katz A, Podder Rakhi, Minko Glenn, Wei Hui, Berriman John, Alfano R R, Gottlieb Paul
Institute for Ultrafast Spectroscopy and Lasers, The City College of New York, NY 10031, USA.
Photochem Photobiol. 2005 Jul-Aug;81(4):879-83. doi: 10.1562/2005-01-14-RA-416.
Native fluorescence spectroscopy (NFS), primarily from tryptophan (trp), was used for in situ investigation of the virus-receptor attachment process in phi6, a lipid-containing bacteriophage from the Cystoviridae family. NFS allowed us to monitor the viral attachment directly to its receptor, which was isolated from the pseudomonad host. Immediately upon mixing, an increase in tryptophan emission intensity was observed followed by a subsequent decrease in emission intensity. The initial increase in emission intensity reflects changes in trp quantum efficiency as the phi6 surface proteins change their conformation as a result of virus attachment to the pilus. The cystovirus spike protein P3 is responsible for receptor recognition and the fluorescence changes observed are likely to be the consequence of its conformational transition at this initial infection stage, providing a kinetic view of this process. The subsequent decrease in trp emission intensity could be due to changes in viral proteins as a result of disassembly of the pili. The technique may have important applications for the dynamic monitoring of additional stages of the virus replication cycle such as assembly, interaction with nucleic acids and maturation. This work expands on a previous demonstration that fluorescence offered a novel tool to detect virus particle interaction with its host cell.
天然荧光光谱法（NFS）主要利用色氨酸（trp），用于对phi6病毒与受体结合过程进行原位研究。phi6是一种来自囊病毒科的含脂噬菌体。NFS使我们能够直接监测病毒与从假单胞菌宿主中分离出的受体的结合情况。混合后立即观察到色氨酸发射强度增加，随后发射强度下降。发射强度的最初增加反映了色氨酸量子效率的变化，这是由于phi6表面蛋白因病毒与菌毛结合而改变其构象所致。囊病毒刺突蛋白P3负责受体识别，观察到的荧光变化可能是其在初始感染阶段构象转变的结果，为该过程提供了动力学视角。色氨酸发射强度随后的下降可能是由于菌毛解体导致病毒蛋白发生变化。该技术可能在动态监测病毒复制周期的其他阶段（如组装、与核酸的相互作用和成熟）方面具有重要应用。这项工作扩展了先前的一项证明，即荧光提供了一种检测病毒颗粒与其宿主细胞相互作用的新工具。
