Investigating ligand-receptor interactions at bilayer surface using electronic absorption spectroscopy and fluorescence resonance energy transfer.
Investigating ligand-receptor interactions at bilayer surface using electronic absorption spectroscopy and fluorescence resonance energy transfer.
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DOI：
10.1021/la300724z
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发表时间：
2012-09-11
期刊：
影响因子：
--
通讯作者：
Kohli P
中科院分区：
文献类型：
--
作者：
Dogra N;Li X;Kohli P
利用电子吸收光谱和荧光共振能量转移（FRET）技术研究了聚二乙炔（PDA）脂质体纳米颗粒双层表面受体和配体之间的相互作用。我们研究了受体和脂质体双层之间的连接模式（共价与非共价）的影响。我们还研究了通过电子吸收和FRET响应的脂质体和分析物之间的大小依赖性相互作用的效果。葡萄糖（受体）分子以共价或非共价方式连接在纳米颗粒的双层上，它们为葡萄糖与E.杆菌E.葡萄糖与配体之间的受体-配体相互作用大肠杆菌表面诱导共轭PDA链上的应力，导致PDA的吸收光谱发生变化（蓝色到红色）。电子吸光度的变化也导致共轭PDA链（受体）和荧光团（磺基罗丹明-101）（供体）连接到双层表面之间的FRET效率的变化。有趣的是，我们没有发现共价和非共价结合的葡萄糖与脂质体在与大肠杆菌相互作用后的UV-Vis和FRET反应有显著差异。杆菌我们将这些结果归因于葡萄糖受体分子与脂质体双层表面的紧密接近，使得在两种情况下诱导的应力相似。我们还发现来自直接激发机制的PDA发射比基于FRET的响应大约2 - 10倍。这些发射信号的差异归因于三个主要原因：E.大肠杆菌和脂质体;分析物和脂质体之间的大小差异;以及相对于磺基罗丹明（SR-101）更高的PDA浓度。我们提出了一个模型来解释我们的实验观察。我们在这里报告的基础研究将有助于提高我们的知识，在分子水平上的软粒子之间的相互作用。
We investigate interactions between receptors and ligands at bilayer surface of polydiacetylene (PDA) liposomal nanoparticles using changes in electronic absorption spectroscopy and Fluorescence Resonance Energy Transfer (FRET). We study the effect of mode of linkage (covalent versus non-covalent) between the receptor and liposome bilayer. We also examine the effect of size dependent interactions between liposome and analyte through electronic absorption and FRET responses. Glucose (receptor) molecules were either covalently or non-covalently attached at the bilayer of nanoparticles, and they provided selectivity for molecular interactions between glucose and glycoprotein ligands of E. coli. The receptor-ligand interactions between glucose and ligand on E. Coli surface induced stress on conjugated PDA chain which resulted in changes (blue to red) in the absorption spectrum of PDA. The changes in electronic absorbance also led to changes in FRET efficiency between conjugated PDA chains (acceptor) and fluorophores (Sulphorhodamine-101) (donor) attached to the bilayer surface. Interestingly, we did not find significant differences in UV-Vis and FRET responses for covalently- and non-covalently-bound glucose to liposomes following their interactions with E. Coli. We attributed these results to close proximity of glucose receptor molecules to the liposome bilayer surface such that induced stress were similar in both the cases. We also found that PDA emission from direct excitation mechanism was ~ 2 - 10 times larger than that of FRET based response. These differences in emission signals were attributed to three major reasons: non-specific interactions between E. Coli and liposomes; size differences between analyte and liposomes; and a much higher PDA concentration with respect to sulpho-rhodamine (SR-101). We have proposed a model to explain our experimental observations. Our fundamental studies reported here will help in enhancing our knowledge regarding interactions involved between soft particles at molecular levels.
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