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宿主多孔蛋白质晶体内客体DNA传输和吸附的表征

Szuhsuan Chen¹, Julius D Stuart², Brian Munsky¹

¹Department of Chemical and Biological Engineering, Colorado State University, Fort Collins, Colorado 80523, United States.

Langmuir : the ACS journal of surfaces and colloids

|December 2, 2024

概括

我们使用计算模型研究了多孔蛋白质晶体中的DNA运输. DNA比以前测量的更快地进入空晶体,揭示了核酸运输动态的洞察力.

科学领域:

生物物理学的生物物理.
材料科学材料科学材料科学
纳米技术纳米技术

背景情况:

通过纳米孔进行核酸运输已经得到了很好的研究.
通过扩展的蛋白质通道进行的传输为长时间的结合相互作用提供了洞察力.
来自Campylobacter jejuni的多孔蛋白质晶体为DNA传输研究提供了一个模型系统.

研究的目的:

为了研究扩展蛋白质通道内的DNA运输和结合.
使用计算方法建模核酸运输.
为了确定双链DNA (dsDNA) 的孔内扩散系数.

主要方法:

使用的Campylobacter jejuni (CJ) 蛋白质的多孔蛋白质晶体.
使用8个基对DNA (8mer) 进行了吸附异温研究.
采用光共聚焦显微镜来追踪DNA加载.
应用了四种有限体积 (FV) 计算模型来分析运输.
利用Fick的第一定律进行扩散系数分析.
在光漂白 (FRAP) 实验后进行光恢复.

主要成果:

CJ蛋白质晶体对8mer DNA (206 ± 30 L/g) 具有很高的平衡吸附常数.
DNA加载主要发生在主要轴性纳米孔沿线.
一个基本的FV模型充分描述了DNA加载动力学.
确定了8mer和15mer dsDNA的体内扩散系数.
将DNA加载到空晶体中的速度比FRAP测量快2个数量级.

结论:

多孔蛋白质晶体适合研究DNA运输和结合.
有限体积法有效地模拟了这些蛋白质通道中的DNA运输.
dsDNA的体内扩散是快速的,特别是在空道.
这项研究为分析核酸运输动态提供了新的策略.

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