Merck Center for Catalysis, Princeton University, Princeton, NJ 08544, USA.
Merck Exploratory Science Center, Merck & Co., Inc., Cambridge, MA 02141, USA.
Science. 2020 Mar 6;367(6482):1091-1097. doi: 10.1126/science.aay4106.
Many disease pathologies can be understood through the elucidation of localized biomolecular networks, or microenvironments. To this end, enzymatic proximity labeling platforms are broadly applied for mapping the wider spatial relationships in subcellular architectures. However, technologies that can map microenvironments with higher precision have long been sought. Here, we describe a microenvironment-mapping platform that exploits photocatalytic carbene generation to selectively identify protein-protein interactions on cell membranes, an approach we term MicroMap (μMap). By using a photocatalyst-antibody conjugate to spatially localize carbene generation, we demonstrate selective labeling of antibody binding targets and their microenvironment protein neighbors. This technique identified the constituent proteins of the programmed-death ligand 1 (PD-L1) microenvironment in live lymphocytes and selectively labeled within an immunosynaptic junction.
许多疾病病理学可以通过阐明局部生物分子网络或微环境来理解。为此，酶邻近标记平台被广泛应用于绘制亚细胞结构中更广泛的空间关系。然而，长期以来一直需要能够更精确地绘制微环境的技术。在这里，我们描述了一种利用光催化卡宾生成来选择性地识别细胞膜上蛋白质-蛋白质相互作用的微环境绘图平台，我们称之为 MicroMap（μMap）。通过使用光催化剂-抗体缀合物在空间上定位卡宾的生成，我们证明了抗体结合靶标及其微环境蛋白邻居的选择性标记。该技术鉴定了活淋巴细胞中程序性死亡配体 1（PD-L1）微环境的组成蛋白，并在免疫突触结内选择性标记。
