Endo-IP and lyso-IP toolkit for endolysosomal profiling of human-induced neurons

• DOI: 10.1073/pnas.2419079121
• Publication Date: 2024-12-05T18:36:47Z
• Authors: Frances V. Hundley; Miguel A. Gonzale...; Lena M. Gottschalk; Aslan N. K. Cook; Jiuchun Zhang; Joao A. Paulo; J. Wade Harper
• Keywords: Synaptogenesis

Plasma membrane protein degradation and recycling are regulated by the endolysosomal system, wherein endocytic vesicles bud from plasma into cytoplasm mature endosomes then degradative lysosomes. As such, system plays a critical role in determining abundance of proteins on cell surface influencing cellular identity function. Highly polarized cells, like neurons, rely for axonal dendritic specialization synaptic compartmentalization. The importance this to neuronal function is reflected prevalence risk variants components several neurodegenerative diseases, ranging Parkinson’s Alzheimer’s disease. Nevertheless, our understanding cargo core machinery neurons limited, part due technical limitations. Here, we develop toolkit capturing EEA1-positive (termed Endo-IP) TMEM192-positive lysosomes Lyso-IP) stem cell-derived induced (iNeurons). We demonstrate its utility revealing landscapes cells cortical-like iNeurons, profiling response potassium-mediated depolarization. Through global cargo, identify hundreds transmembrane proteins, including neurogenesis as well with predicted SNX17 or SNX27 recognition motifs. By contrast, parallel lysosome reveals simpler repertoire, reflecting temporally controlled many targets. This will facilitate mechanistic interrogation found factors