CAT-S proteomic data (searched result)

In situ profiling of subcellular proteomic networks in primary and living systems, such as primary cells from native tissues or clinic samples, is crucial for the understanding of life processes and diseases, yet challenging for the current proximity labeling methods (e.g., BioID, APEX) due to their necessity of genetic engineering. Here we report CAT-S, an up-to-date bioorthogonal photocatalytic chemistry-enabled proximity labeling method, that expands proximity labeling to a wide range of primary living samples for in situ profiling of subcellular proteomes. Powered by the newly introduced thioQM labeling warhead and targeted bioorthogonal photocatalytic decaging chemistry, CAT-S enables labeling of mitochondrial proteins in living cells with high efficiency and specificity (up to 87%). We applied CAT-S to distinct cell cultures, mouse tissues as well as primary T cells from human blood, portraying the native-state mitochondrial proteomic characteristics, and unveiled a set of hidden mitochondrial proteins in human proteome. Furthermore, CAT-S allowed quantitative analysis of the in situ proteomic perturbations on dysfunctional tissue samples, exampled by diabetic mouse kidneys, and revealed the alterations of lipid metabolism machinery that drive the disease progression. Given the advantages of non-genetic operation, generality, efficiency as well as spatiotemporal resolution, CAT-S may open new avenues as a proximity labeling strategy for in situ investigation of subcellular proteomic landscape of primary living samples that are otherwise inaccessible.

针对原代活体系统（例如源自天然组织或临床样本的原代细胞）的亚细胞蛋白质组网络原位谱分析，对于解析生命过程与疾病机制至关重要，但现有邻近标记技术（如BioID、APEX）因依赖基因工程操作，难以适配此类研究。本研究报道了CAT-S：一种基于新一代生物正交光催化化学的邻近标记技术，该技术将邻近标记的适用范围拓展至多种原代活体样本，用于亚细胞蛋白质组的原位谱分析。依托全新开发的硫代醌甲基化物（thioQM）标记弹头与靶向生物正交光催化脱笼化学，CAT-S可在活细胞中高效且特异性地标记线粒体蛋白质，标记效率最高可达87%。研究团队将CAT-S应用于不同细胞培养体系、小鼠组织以及人血液来源的原代T细胞，表征了天然状态下的线粒体蛋白质组特征，并揭示了人类蛋白质组中一组隐匿的线粒体蛋白质。此外，CAT-S可实现功能异常组织样本的原位蛋白质组扰动定量分析，以糖尿病小鼠肾脏为例，该技术揭示了驱动疾病进展的脂质代谢机制的改变。鉴于CAT-S具备非基因操作、普适性强、高效性与优良时空分辨率等优势，作为一种邻近标记策略，它可为原代活体样本的亚细胞蛋白质组图谱原位研究开辟全新路径，而此类样本此前往往难以开展相关研究。