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Insect Expression> | … To obtain GST-FIP200 and GST-FIP200-GFP constructs the insect codon-optimized FIP200 gene was purchased from GenScript and cloned with the respective tags into pGB-02-03 via the Gibson assembly method by the Vienna BioCenter Core Facilities (VBCF) Protech Facility… | Get A Quote |
The autophagic degradation of misfolded and ubiquitinated proteins is important for cellular homeostasis. In this process, which is governed by cargo receptors, ubiquitinated proteins are condensed into larger structures and subsequently become targets for the autophagy machinery. Here we employ in vitro reconstitution and cell biology to define the roles of the human cargo receptors p62/SQSTM1, NBR1 and TAX1BP1 in the selective autophagy of ubiquitinated substrates. We show that p62 is the major driver of ubiquitin condensate formation. NBR1 promotes condensate formation by equipping the p62-NBR1 heterooligomeric complex with a high-affinity UBA domain. Additionally, NBR1 recruits TAX1BP1 to the ubiquitin cond... More