首页    期刊浏览 2025年06月11日 星期三
登录注册

文章基本信息

  • 标题:In vitro reconstitution of calcium-dependent recruitment of the human ESCRT machinery in lysosomal membrane repair
  • 本地全文:下载
  • 作者:Sankalp Shukla ; Kevin P. Larsen ; Chenxi Ou
  • 期刊名称:Proceedings of the National Academy of Sciences
  • 印刷版ISSN:0027-8424
  • 电子版ISSN:1091-6490
  • 出版年度:2022
  • 卷号:119
  • 期号:35
  • DOI:10.1073/pnas.2205590119
  • 语种:English
  • 出版社:The National Academy of Sciences of the United States of America
  • 摘要:Significance One of the ways by which protein aggregates can propagate and lead to the progression of a neurodegenerative disease is by damaging the membrane that is destined to degrade these misfolded aggregates. The ESCRT machinery has been implicated in sealing these damaged membranes, and the nature of the membrane recruitment trigger signal for this machinery is a major open question. Here, we show in vitro that ALG-2 alone is sufficient to bring the ESCRT machinery to membranes in a Ca 2+-dependent manner. The endosomal sorting complex required for transport (ESCRT) machinery is centrally involved in the repair of damage to both the plasma and lysosome membranes. ESCRT recruitment to sites of damage occurs on a fast time scale, and Ca 2+ has been proposed to play a key signaling role in the process. Here, we show that the Ca 2+-binding regulatory protein ALG-2 binds directly to negatively charged membranes in a Ca 2+-dependent manner. Next, by monitoring the colocalization of ALIX with ALG-2 on negatively charged membranes, we show that ALG-2 recruits ALIX to the membrane. Furthermore, we show that ALIX recruitment to the membrane orchestrates the downstream assembly of late-acting CHMP4B, CHMP3, and CHMP2A subunits along with the AAA + ATPase VPS4B. Finally, we show that ALG-2 can also recruit the ESCRT-III machinery to the membrane via the canonical ESCRT-I/II pathway. Our reconstitution experiments delineate the minimal sets of components needed to assemble the entire membrane repair machinery and open an avenue for the mechanistic understanding of endolysosomal membrane repair.
  • 关键词:enmembrane biologymembrane remodelingmembrane repairin vitro reconstitutionneurodegeneration
国家哲学社会科学文献中心版权所有