Alkyne tyramide is a clickable ascorbate peroxidase 2 (APEX2) probe with water solubility and cell wall permeability that significantly improves APEX labeling efficiency in intact yeast cells and also AIDS in the identification of APEX labeling sites that can clearly assign the membrane topology of mitochondrial proteins[1].
The engineered ascorbate peroxidase (APEX) is a powerful tool for the proximity-dependent labeling of proteins and RNAs in live cells. Alkyne tyramide was identified as a clickable APEX substrate that enables proteomic analysis in the mitochondrial matrix of intact yeast cells with extremely high specificity (94%) and higher coverage than the traditional APEX2 substrate biotin-phenol (BP). Alkyne tyramide labeling is dependent on APEX2 expression and the presence of H2O2, and APEX2-mediated alkyne tyramide labeling of yeast mitochondria is more efficient when the probe concentration is greater than 1mM[1].
References:
[1] Li Y, Tian C, Liu K, et al. A clickable APEX probe for proximity-dependent proteomic profiling in yeast[J]. Cell Chemical Biology, 2020, 27(7): 858-865. e8.
Alkyne tyramide是一种可点击的抗坏血酸过氧化物酶2（APEX2）探针，具有水溶性和细胞壁渗透性，可显著提高完整酵母细胞中的APEX标记效率，还有助于识别APEX标记位点，从而可以明确分配线粒体蛋白的膜拓扑结构[1]。
工程化抗坏血酸过氧化物酶（APEX）是活细胞中蛋白质和RNA邻近依赖性标记的强大工具。Alkyne tyramide被鉴定为可点击的APEX底物，该底物能够以极高的特异性（94%）在完整酵母细胞的线粒体基质中进行蛋白质组学分析，并且比传统的APEX2底物生物素-苯酚（BP）具有更高的覆盖率。Alkyne tyramide标记取决于APEX2表达和H2O2的存在，并且当探针浓度大于1mM时，APEX2介导的酵母线粒体Alkyne tyramide标记更加有效[1]。