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Measurements of Ion Binding to Lipid-Hosted Ionophores by Affinity Chromatography.
Langmuir ( IF 3.7 ) Pub Date : 2019-06-24 00:00:00 , DOI: 10.1021/acs.langmuir.9b01301
Eric E Ross 1 , Bridget Hoag 1 , Ian Joslin 1 , Taylor Johnston 1
Affiliation  

The binding affinity between antibiotic ionophores and alkali ions within supported lipid bilayers was evaluated using affinity chromatography. We used zonal elution and frontal analysis methods in nanovolume liquid chromatography to characterize the binding selectivity of the carrier and channel ionophores valinomycin and gramicidin A within different phosphatidylcholine bilayers. Distinct binding sensitivity to the lipid phase, both in affinity and selectivity, is observed for valinomycin, whereas gramicidin is less sensitive to changes in a membrane environment, behavior that is consistent with ion binding occurring within the interior of an established channel. There is good agreement between the chromatographic retention and the reported binding selectivity measured by other techniques. Surface potential near the binding site affects ion retention and the apparent association binding constants, but not the binding selectivity or enthalpy measurements. A model accounting for the surface potential contributions of retained ions during frontal analyses yields values close to intrinsic binding constants for gramicidin A (KA for K+ between 70 and 120 M–1) using reasonable estimates of the initial potential that is postulated to arise from the underlying silica.

中文翻译:

通过亲和色谱法测量离子与脂质基离子载体的结合。

使用亲和色谱法评估了支持的脂质双层中抗生素离子载体与碱金属离子之间的结合亲和力。我们在纳米体积液相色谱中使用了区域洗脱和前沿分析方法来表征不同磷脂酰胆碱双层中载体和通道离子载体缬氨霉素和短杆菌肽A的结合选择性。对于缬氨霉素,在亲和力和选择性上都观察到了对脂质相的不同结合敏感性,而短杆菌肽对膜环境的变化较不敏感,其行为与在已建立通道内部发生的离子结合相一致。在色谱保留度和通过其他技术测得的报道的结合选择性之间有很好的一致性。结合位点附近的表面电势影响离子保留和表观缔合结合常数,但不影响结合选择性或焓测量。在正面分析过程中考虑保留离子的表面电势贡献的模型所产生的值接近于短杆菌肽A的固有结合常数(ķ对于K + 70和120米之间-1使用被假定从底层二氧化硅产生的初始电位的合理估计)。
更新日期:2019-06-24
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