category
bioRxiv
date
Mar 1, 2026
slug
status
Published
summary
创新性提出半舱FRAP实验方法用于螺旋细菌蛋白质扩散研究,建立扩散系数与荧光恢复时间的定量关系模型,并首次在螺旋菌中实现mNeonGreen荧光蛋白扩散系数的定量测量。
tags
蛋白质组学
type
Post

📄 原文题目

Experimental and simulated FRAP for the quantitative determination of protein diffusion in helical cells

🔗 原文链接

💡 AI 核心解读

创新性提出半舱FRAP实验方法用于螺旋细菌蛋白质扩散研究,建立扩散系数与荧光恢复时间的定量关系模型,并首次在螺旋菌中实现mNeonGreen荧光蛋白扩散系数的定量测量。

📝 英文原版摘要

Fluorescence recovery after photobleaching (FRAP) is widely used to characterize diffusion in cells, but quantitative interpretation of the data in small prokaryotes requires explicitly accounting for cell geometry. While this has been successfully achieved for spherical and rod-shaped bacteria, analytical approaches developed in these cases are not directly applicable to cells with more complex morphologies. Here, we explore the application of FRAP to helical bacteria using simulations. We show that half-compartment FRAP experiments, where one-half of the cell is photobleached, provide a robust means of characterizing fast protein diffusion. To help with the practical implementation of this technique, we established the relationship between the diffusion coefficient and characteristic fluorescence recovery time as a function of cell length and helical parameters, and for two different ways of estimating the recovery time. As a first application, we report measurements of the diffusion coefficient of the fluorescent protein, mNeonGreen, in the helical bacterium textit{Paramagnetospirillum magneticum} AMB-1. We find it to be D = 4.9 +/- 2.2 m^2/s in isosmotic conditions, not significantly different from the value measured in Escherichia coli. Although developed for helical bacteria, including spirilla, spirochetes, and vibrios, our framework can readily be extended to cells or compartments with other geometries.
细菌蛋白酶体激活剂Bpa的结构异质性及底物结合机制构象变化在TcmN芳香酶/环化酶聚酮类生物合成中的作用
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