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Proteins, Expression, Isolation and Analysis> | … The supernatants were first purified with Ni-NTA resin (GenScript), followed by purification on a Superdex 200 increase 10/300 column (SD200) (GE healthcare). All proteins were stored in storage buffer (50 mM Tris-HCl pH 7.4, 150 mM NaCl) at −80 C. All protein purification … | Get A Quote |
Liquid-liquid phase separation promotes the formation of membraneless condensates that mediate diverse cellular functions, including autophagy of misfolded proteins. However, how phase separation participates in autophagy of dysfunctional mitochondria (mitophagy) remains obscure. We previously discovered that nuclear receptor Nur77 (also called TR3, NGFI-B, or NR4A1) translocates from the nucleus to mitochondria to mediate celastrol-induced mitophagy through interaction with p62/SQSTM1. Here, we show that the ubiquitinated mitochondrial Nur77 forms membraneless condensates capable of sequestrating damaged mitochondria by interacting with the UBA domain of p62/SQSTM1. However, tethering clustered mitochondria to... More