{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,6]],"date-time":"2026-03-06T01:03:30Z","timestamp":1772759010093,"version":"3.50.1"},"reference-count":37,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2002,12,3]],"date-time":"2002-12-03T00:00:00Z","timestamp":1038873600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["FEBS Letters"],"published-print":{"date-parts":[[2002,12,18]]},"abstract":"<jats:p>Autophagosome formation is a central event in macroautophagy. The Apg12\u2013Apg5 conjugate, which is essential in this process, is generated by a ubiquitin\u2010like protein conjugation system. In yeast, Apg12, following activation by the E1\u2010like Apg7, forms a thioester with Apg10 (E2\u2010like). Apg12 is finally conjugated to Apg5 via an isopeptide bond. The possible requirement of an E3\u2010like protein for the conjugation, however, has not yet been confirmed. The Apg12 system is conserved among eukaryotes, although a mammalian counterpart of Apg10 has not yet been identified. Here, we report the identification and characterization of the mouse Apg10 ortholog. A yeast two\u2010hybrid screen using the mouse Apg5 (mApg5) as bait identified a novel protein with 19% identity to yeast Apg10. We designated this protein mouse Apg10 (mApg10). We demonstrated by a modified yeast two\u2010hybrid assay that mApg10 mediates the conjugation of mApg12 and mApg5. The in vivo interaction of mApg12 with mApg10 in HeLa cells suggests that mApg10 is an Apg12\u2010conjugating enzyme, likely serving as an Apg5\u2010recognition molecule in the Apg12 system. This novel two\u2010hybrid method, which we have named \u2018conjugation\u2010mediated yeast two\u2010hybrid\u2019, proves to be a simple and useful technique with which to analyze protein\u2013protein conjugation.<\/jats:p>","DOI":"10.1016\/s0014-5793(02)03739-0","type":"journal-article","created":{"date-parts":[[2002,12,27]],"date-time":"2002-12-27T16:51:48Z","timestamp":1041007908000},"page":"450-454","source":"Crossref","is-referenced-by-count":42,"title":["Mouse Apg10 as an Apg12\u2010conjugating enzyme: analysis by the conjugation\u2010mediated yeast two\u2010hybrid 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