Surfaceomic technologies and antibodies to probe cell surface proteomes and their interactomes at unprecedented small scale and high-resolution

标题：探测细胞表面蛋白质组及其相互作用的表面组学技术和抗体 史无前例的小尺度和高分辨率 项目总结 细胞表面体是允许细胞感知和响应环境变化的主要中枢， 然而，在估计的4000-5000种质膜和分泌的蛋白质中，只有一小部分具有功能。 特色化的。我们对表面体的协调调节了解得更少，特别是如何 膜蛋白的组成、复合体、分布和功能被改变以共同调节 细胞表型的变化。我们的长期目标是系统地了解细胞是如何重塑的 它们的膜蛋白质组(表面蛋白质组)在健康和疾病中的作用，并开发探测和 调节这些过程。在过去五年里，在R35 Mira的支持下，我们广泛地 表征了癌基因转化引起的表面体变化，从而发现了新的 生物标记物、抗癌药物靶标和高特异性抗体的产生。在未来五年，我们将 开发新的表面组学技术用于小规模的细胞群体分析，特别是在 使用IPSC衍生模型和患者衍生样本的神经退行性变。我们将定义细胞表面 使用Dexter Energy实现的高分辨率邻近标记方法的蛋白质复合体和相互作用 转移(Det)。我们计划审问 I、II、III中EGFR家族受体的相互作用组 癌症与T细胞中的细胞因子受体 红外 激活。我们相信这项技术 小规模的表面组学和互动组学将 显著提高我们绘制细胞图的能力 疾病中的表面及其内部 从癌症，免疫学，神经学， 还有更多。我们制作了重组图1。表面组学技术发展概述。 使用针对100种细胞表面蛋白的抗体 噬菌体、酵母和哺乳动物的展示，我们期待着在理解表面组方面的进一步努力。 将促进高选择性抗体的发展，这些抗体可以探测和调节细胞状态。我们开始了 这一愿景在我们的第一个R35拨款中通过定义由癌症中特定癌基因诱导的表面体来实现。接下来，我们 将极大地将表面组学科学扩展到更小规模的特化和原代细胞，以及更多 更高分辨率的信号相互作用分析。

Title: Surfaceomic technologies and antibodies to probe cell surface proteomes and their interactomes at unprecedented small scale and high-resolution Project summary The cell surfaceome is the primary hub that allows cells to sense and respond to changes in their environment, yet only a minority of the estimated 4000-5000 plasma membrane and secreted proteins have been functionally characterized. We understand even less about coordinate regulation of the surfaceome, and specifically how the composition, complexes, distribution, and function of membrane proteins are altered to collectively mediate changes to cellular phenotypes. Our long-range goal is to systematically understand how cells remodel their membrane proteome (surfaceome) in health and disease, and to develop antibodies that probe and modulate these processes. With the support of the R35 MIRA in the last five years, we have extensively characterized the surfaceome changes induced by oncogene transformation leading to discovery of new biomarkers, cancer drug targets, and generation of highly specific antibodies. In the next five years, we will develop new surfaceome technologies for small scale analysis of cell populations, particularly in neurodegeneration using iPSC derived models and patient-derived samples. We will define the cell surface protein complexes and interactomes using a high-resolution proximity labeling method enabled by Dexter Energy Transfer (DET). We plan to interrogate the interactome of EGFR family receptors in i ii iii cancer, and cytokine receptors in T-cell Ir activation. We believe the technologies for small scale surfaceomics and interactomics will dramatically improve our abilities to map cell surfaces and their intreactomes in diseases ranging from cancer, immunology, neurology, and more. We have produced recombinant Figure 1. Overview of development of surfaceomics technologies. antibodies to 100s of cell surface proteins using phage, yeast, and mammalian display, and we anticipate the expanded efforts in understanding the surfaceome will advance the development of highly selective antibodies that probe and modulate cellular states. We began this vision in our first R35 grant by defining the surfaceomes induced by specific oncogenes in cancer. Next, we will greatly expand surfaceomics science to much smaller scales of specialized and primary cells, and much higher resolution analysis of signaling interactomes.