Elucidating the cellular determinants of targeted membrane protein degradation by lysosome-targeting chimeras

• 成果类型: Article
• 署名作者: Ahn, Green; Riley, Nicholas M.; Kamber, Roarke A.; Wisnovsky, Simon; von Hase, Salvador Moncayo; Bassik, Michael C.; Banik, Steven M.; Bertozzi, Carolyn R.
• 署名单位: Stanford University; Stanford University; Stanford University; University of British Columbia; Stanford University; Howard Hughes Medical Institute
• 刊物名称: SCIENCE
• ISSN/ISSBN: 0036-13888
• DOI: 10.1126/science.adf6249
• 发表日期: 2023-10-20
• 关键词: factor-ii receptor; mannose 6-phosphate receptors; intracellular trafficking; computational platform; identification; cullin; retromer; delivery; binding; ubiquitination

摘要

Targeted protein degradation can provide advantages over inhibition approaches in the development of therapeutic strategies. Lysosome-targeting chimeras (LYTACs) harness receptors, such as the cation-independent mannose 6-phosphate receptor (CI-M6PR), to direct extracellular proteins to lysosomes. In this work, we used a genome-wide CRISPR knockout approach to identify modulators of LYTAC-mediated membrane protein degradation in human cells. We found that disrupting retromer genes improved target degradation by reducing LYTAC recycling to the plasma membrane. Neddylated cullin-3 facilitated LYTAC-complex lysosomal maturation and was a predictive marker for LYTAC efficacy. A substantial fraction of cell surface CI-M6PR remains occupied by endogenous M6P-modified glycoproteins. Thus, inhibition of M6P biosynthesis increased the internalization of LYTAC-target complexes. Our findings inform design strategies for next-generation LYTACs and elucidate aspects of cell surface receptor occupancy and trafficking.