Autophagy Related Protein 10 (ATG10)

The Atg10-ATG12 interaction required the C-terminal glycine of ATG12. Atg10 with a cys161-to-ser (C161S) mutation formed a stable intermediate with ATG12 in the presence of ATG7, suggesting that C161 is the active-site cysteine for the E2-like reaction of ATG12. Furthermore, overexpression of Atg10 facilitated modification of a soluble form of MAP-LC3 to a membrane-bound form in an ATG7-dependent manner. Immunoprecipitation analysis of transfected human embryonic kidney cells and yeast 2-hybrid analysis showed that mouse Atg10 interacted with MAP-LC3 directly in an ATG7-independent manner. Atg10 with the C161S mutation did not form an intermediate with MAP-LC3, even in the presence of ATG7, indicating that MAP-LC3 is not a substrate of Atg10.