通过吸附离子的饱和来确定细菌表面电荷密度,bioRxiv - Biophysics

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通过吸附离子的饱和来确定细菌表面电荷密度
bioRxiv - Biophysics Pub Date : 2020-10-01 , DOI: 10.1101/2020.09.29.318840
M.J. Wilhelm , M. Sharifian Gh. , C.M. Chang , T. Wu , Y. Li , J. Ma , H.L. Dai

细菌表面电荷是细胞界面生理的关键特征，它会影响细胞与局部环境的相互作用。需要一种能够量化细菌表面电荷的直接，灵敏，准确的实验技术，以更好地理解分子生理学对环境变化的适应性。我们在这里介绍二次谐波光散射（SHS）的方法，该方法能够检测作为反电荷吸附在细菌外表面的分子离子的数量，从而提供表面电荷的量度。在第一个演示中，我们检测到静电吸附在革兰氏阳性和革兰氏阴性细菌代表性菌株表面的小分子阳离子，孔雀石绿。出奇，SHS推断通过不同细胞超微结构的分子转运速率几乎相同。然而，两种细菌的外表面上的吸附饱和密度显示出特征上不同。革兰氏阴性大肠杆菌的脂多糖包被的外表面的负电荷密度（8.7±1.7 nm-2）被推论为鼠李糖乳杆菌的蛋白质表面层（1.2±0.2）的负电荷密度的七倍。 nm-2）。提出了SHS推断细菌表面电荷密度用于革兰氏型分化的可行性。革兰氏阴性大肠杆菌的脂多糖包被的外表面的负电荷密度（8.7±1.7 nm-2）被推论为鼠李糖乳杆菌的蛋白质表面层（1.2±0.2）的负电荷密度的七倍。 nm-2）。提出了SHS推断细菌表面电荷密度用于革兰氏型分化的可行性。革兰氏阴性大肠杆菌的脂多糖包被的外表面的负电荷密度（8.7±1.7 nm-2）被推论为鼠李糖乳杆菌的蛋白质表面层（1.2±0.2）的负电荷密度的七倍。 nm-2）。提出了SHS推断细菌表面电荷密度用于革兰氏型分化的可行性。

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Determination of Bacterial Surface Charge Density Via Saturation of Adsorbed Ions

Bacterial surface charge is a critical characteristic of the cell's interfacial physiology that influences how the cell interacts with the local environment. A direct, sensitive, and accurate experimental technique capable of quantifying bacterial surface charge is needed to better understand molecular adaptations in interfacial physiology in response to environmental changes. We introduce here the method of second harmonic light scattering (SHS) which is capable of detecting the number of molecular ions adsorbed as counter charges on the exterior bacterial surface, thereby providing a measure of the surface charge. In this first demonstration, we detect the small molecular cation, malachite green, electrostatically adsorbed on the surface of representative strains of Gram-positive and Gram-negative bacteria. Surprisingly, the SHS deduced molecular transport rates through the different cellular ultra-structures are revealed to be nearly identical. However, the adsorption saturation densities on the exterior surfaces of the two bacteria were shown to be characteristically distinct. The negative charge density of the lipopolysaccharide coated outer surface of Gram-negative E. coli (8.7±1.7 nm-2) was deduced to be seven times larger than that of the protein surface layer of Gram-positive L. rhamnosus (1.2±0.2 nm-2). The feasibility of SHS deduced bacterial surface charge density for Gram-type differentiation is presented.

更新日期：2020-10-02

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