Department of Microbiology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.
Department of Bacteriology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Probiotics Antimicrob Proteins. 2020 Jun;12(2):697-704. doi: 10.1007/s12602-019-09573-2.
This study proposed to investigate the effect of azurin on the major stages of pathogenesis (adhesion and invasion) of intestinal bacterial pathogens (Salmonella spp. and Escherichia coli) and epithelial pathogens (Staphylococcus aureus and Pseudomonas aeruginosa) on the human colorectal adenocarcinoma (Caco-2) cell line. Azurin protein was produced by cloning the azurin gene into pET21a and heterologous expression in E. coli BL21. The protein was purified using affinity chromatography and confirmed by Western blotting. The purified protein was evaluated by three experiments of adhesion and invasion assays, including exclusion, competition, and replacement. Azurin was observed to significantly inhibit the attachment and invasion of S. aureus, Salmonella spp., and E. coli, while no such inhibitory effects were observed on P. aeruginosa. In fact, the protein increased the adhesion of P. aeruginosa to the cell. In conclusion, our study proposes that azurin is a potential prophylactic or preventive helper candidate to inhibit the attachment and invasion of pathogenic bacteria to host cells and reduce the progression of the infection process. Our study also reveals the involvement of azurin in bacteria-host cell interactions, providing novel and important insights toward the elucidation of its biological function in this field. Thus, this study provides new opportunities to use azurin as an adjunct therapy against critical stages of infection by a wide range of pathogenic bacteria.
本研究旨在探讨蓝蛋白对肠道细菌病原体（沙门氏菌和大肠杆菌）和上皮病原体（金黄色葡萄球菌和铜绿假单胞菌）在人结直肠腺癌细胞系（Caco-2）上发病主要阶段（黏附和侵袭）的影响。蓝蛋白蛋白通过将蓝蛋白基因克隆到 pET21a 中并在大肠杆菌 BL21 中异源表达来生产。使用亲和层析法纯化蛋白质，并通过 Western blot 进行确认。通过黏附和侵袭实验的三个实验（排除、竞争和替代）评估纯化的蛋白质。蓝蛋白显著抑制金黄色葡萄球菌、沙门氏菌和大肠杆菌的附着和侵袭，而对铜绿假单胞菌则没有这种抑制作用。事实上，该蛋白增加了铜绿假单胞菌对细胞的黏附。总之，我们的研究表明蓝蛋白是一种潜在的预防或预防性辅助候选物，可抑制致病菌对宿主细胞的附着和侵袭，从而减缓感染过程的进展。我们的研究还揭示了蓝蛋白在细菌-宿主细胞相互作用中的作用，为阐明其在该领域的生物学功能提供了新的和重要的见解。因此，这项研究为使用蓝蛋白作为辅助治疗提供了新的机会，以对抗各种致病菌感染的关键阶段。
