Department of Molecular Microbiology and Biotechnology, George S. Wise Faculty of Life Sciences, Tel Aviv University, Tel Aviv 6997801, Israel.
Department of Materials Science and Engineering, Iby and Aladar Fleischman Faculty of Engineering, Tel Aviv University, Tel Aviv 6997801, Israel.
Molecules. 2018 May 26;23(6):1273. doi: 10.3390/molecules23061273.
The amino acid tyrosine forms cytotoxic amyloid-like fibrils by molecular self-assembly. However, the production of antibodies towards tyrosine assemblies, reflecting their presentation to the immune system, was not demonstrated yet. Here, we describe the production of antibodies that specifically recognize tyrosine in its fibrillated form. The antibodies were demonstrated to specifically bind self-assembled tyrosine, in contrast to its non-aggregated form or disintegrated fibrils. The antibodies could be used for immunostaining of tyrosine fibrils in cultured cells. Furthermore, confocal microscopy allowed a demonstration of the intracellular presence of the metabolite amyloids in a neuroblastoma cell model. Finally, pre-incubation of tyrosine fibrils with the antibodies resulted in significant reduction in their cytotoxicity. Taken together, we provide an experimental proof for the immunogenicity of tyrosine amyloid fibrillary assemblies. These specific antibodies against tyrosine structures could be further used as a research tool to study the dynamics, toxicity and cellular localization of the assemblies.
氨基酸酪氨酸通过分子自组装形成细胞毒性类似淀粉样的纤维。然而，目前尚未证明针对酪氨酸组装体的抗体的产生，反映了它们在免疫系统中的呈现。在这里，我们描述了产生特异性识别纤维状酪氨酸的抗体。与非聚集形式或解聚纤维相比，这些抗体被证明能够特异性结合自组装的酪氨酸。该抗体可用于免疫染色培养细胞中的酪氨酸纤维。此外，共聚焦显微镜允许在神经母细胞瘤细胞模型中证明细胞内存在代谢物淀粉样蛋白。最后，用抗体预先孵育酪氨酸纤维可显著降低其细胞毒性。总之，我们为酪氨酸淀粉样纤维状组装物的免疫原性提供了实验证据。这些针对酪氨酸结构的特异性抗体可以进一步用作研究工具，以研究组装体的动力学、毒性和细胞定位。
