Intracellular cartography of the human proteome.

Understanding how the proteome is spatially organized within cells remains a central challenge in cell biology. Here, I will present integrative methods for mapping the intracellular architecture of the human proteome at scale. We combine CRISPR-based endogenous tagging, fluorescence live-cell imaging, and quantitative mass spectrometry to generate high-resolution spatial and interaction proteomic maps of human cells. Using graph-based analysis and machine learning, we delineate dynamic protein networks, uncover functional microdomains, and systematically annotate subcellular localization. I will highlight recent advances from our OpenCell and organelle proteomics efforts, including a global map of subcellular organization derived from native immunoprecipitation of tagged organelles. Our work highlights that many cellular responses to perturbation are governed by spatial remodeling rather than by abundance changes. Together, these approaches build a quantitative foundation for charting the cell as a dynamic, spatially organized proteome.