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Interaction mechanism of Mycobacterium tuberculosis GroEL2 protein with macrophage Lectin-like, oxidized low-density lipoprotein receptor-1: An integrated computational and experimental study
Biochimica et Biophysica Acta (BBA) - General Subjects ( IF 3 ) Pub Date : 2020-10-06 , DOI: 10.1016/j.bbagen.2020.129758
Vivek Vinod , Anju Choorakottayil Pushkaran , Anil Kumar , Chethampadi Gopi Mohan , Raja Biswas

Background:

Bacterial surface proteins act as potential adhesins or invasins. The GroEL is a signal peptide-free surface expressed protein that aids adhesion in Escherichia coli by binding to LOX-1 receptor of the host cells. Mycobacterium tuberculosis (Mtb) expresses GroEL2 protein, having high level sequence identity with E. coli GroEL. This study investigates the interaction mechanism of GroEL2 protein of Mtb with LOX-1 of macrophages using integrated computational and experimental approach.

Methods:

Mtb GroEL2 protein was purified as histidine tagged protein using Ni-NTA chromatography. Confocal and scanning electron microscopies were used to study the uptake of GroEL2 coated fluorescent latex beads through the LOX-1 receptor in RAW264.7 macrophage cell line. Docking studies were performed to understand the interaction between the GroEL2 and LOX-1 proteins. Polyinosinic acid (PIA) was used as a LOX-1 inhibitor in both in silico and in vitro experiments.

Results:

GroEL2 protein coating enhances uptake of latex beads into macrophages through LOX-1 receptor. LOX-1 inhibitor PIA decreased the uptake of GroEL2 coated latex beads. GroEL2 interacts with the key ligand binding regions of the LOX-1 receptor, such as the basic spine and the saddle hydrophobic patch. PIA molecule destabilized the LOX-1-GroEL2 docked complex.

Conclusion:

Surface associated GroEL2 protein of Mtb is a potential ligand for macrophage LOX-1 receptor. Interaction between GroEL2 and LOX-1 receptor may be utilized by Mtb to gain its intracellular access.

General Significance:

Surface associated GroEL2 of Mtb may bind to the macrophage LOX-1 receptor, enabling the internalization of the bacteria and progression of the infection.



中文翻译:

结核分枝杆菌GroEL2蛋白与巨噬细胞凝集素样氧化低密度脂蛋白受体-1的相互作用机制:综合计算和实验研究

背景:

细菌表面蛋白可作为潜在的粘附素或入侵素。GroEL是一种无信号肽的表面表达蛋白,通过与宿主细胞的LOX-1受体结合,有助于在大肠杆菌中粘附。结核分枝杆菌Mtb)表达与大肠杆菌GroEL具有高度序列同一性的GroEL2蛋白。本研究采用综合的计算和实验方法研究了Mtb的GroEL2蛋白与巨噬细胞LOX-1的相互作用机理。

方法:

使用Ni-NTA色谱法将Mtb GroEL2蛋白纯化为组氨酸标记的蛋白。共聚焦和扫描电子显微镜用于研究通过RAW264.7巨噬细胞系中的LOX-1受体摄取GroEL2涂覆的荧光胶乳珠。进行了对接研究以了解GroEL2和LOX-1蛋白之间的相互作用。聚肌苷酸(PIA)在计算机体外实验中均用作LOX-1抑制剂。

结果:

GroEL2蛋白涂层可通过LOX-1受体增强乳胶珠对巨噬细胞的吸收。LOX-1抑制剂PIA降低了GroEL2包被的乳胶珠的摄取。GroEL2与LOX-1受体的关键配体结合区相互作用,例如基本脊柱和鞍形疏水膜。PIA分子破坏了LOX-1-GroEL2对接复合物的稳定性。

结论:

Mtb与表面相关的GroEL2蛋白是巨噬细胞LOX-1受体的潜在配体。Mtb可利用GroEL2与LOX-1受体之间的相互作用来获得其细胞内通路。

一般意义:

Mtb的与表面相关的GroEL2可能与巨噬细胞LOX-1受体结合,从而使细菌内化和感染进程。

更新日期:2020-10-16
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