title: Crystallographic Characterization of ATG Proteins and Their Interacting Partners.
abstract: Autophagosome formation and specific substrate recruitment during autophagy require ligation of the ubiquitin-like protein (UBL) Atg8 to the head group of the lipid phosphatidylethanolamine. Atg8 lipidation is mediated by distinctive UBL cascades involving autophagy-specific E1, E2, and E3 enzymes that differ substantially in sequence from components of other UBL conjugation cascades. Structural studies are important for elucidating the roles of Atg proteins that regulate multiple steps involved in autophagy. This chapter describes methods to prepare and crystallize selected proteins and complexes involved in autophagy UBL conjugation pathways, as a guide for strategies for structural and biochemical characterization of Atg proteins.
authors.affiliation: St. Jude Children's Research Hospital, Memphis, TN, United States.
authors.forename: Y
authors.initials: Y
authors.lastname: Qiu
authors.affiliation: St. Jude Children's Research Hospital, Memphis, TN, United States; University of Tennessee Health Sciences Center, Memphis, TN, United States.
authors.forename: Y
authors.initials: Y
authors.lastname: Zheng
authors.affiliation: St. Jude Children's Research Hospital, Memphis, TN, United States.
authors.forename: A M
authors.initials: AM
authors.lastname: Taherbhoy
authors.affiliation: St. Jude Children's Research Hospital, Memphis, TN, United States.
authors.forename: S E
authors.initials: SE
authors.lastname: Kaiser
authors.affiliation: St. Jude Children's Research Hospital, Memphis, TN, United States; University of Tennessee Health Sciences Center, Memphis, TN, United States; Howard Hughes Medical Institute, St. Jude Children's Research Hospital, Memphis, TN, United States. Electronic address: brenda.schulman@stjude.org.
authors.forename: B A
authors.initials: BA
authors.lastname: Schulman
mesh_terms: D001343:Autophagy; D000071187:Autophagy-Related Protein 5 / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D000071193:Autophagy-Related Protein 7 / Q000737:chemistry / Q000378:metabolism; D000071183:Autophagy-Related Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D003460:Crystallization / Q000379:methods; D003461:Crystallography / Q000379:methods*; D006801:Humans; D009154:Mutation; D000072417:Protein Domains; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000378:metabolism; D044763:Ubiquitin-Conjugating Enzymes / Q000737:chemistry / Q000378:metabolism; D044767:Ubiquitin-Protein Ligases
chemical_list: C502778:ATG5 protein, human; C114830:ATG7 protein, S cerevisiae; D000071187:Autophagy-Related Protein 5; D000071183:Autophagy-Related Proteins; D029701:Saccharomyces cerevisiae Proteins; C557619:ATG10 protein, S cerevisiae; D044763:Ubiquitin-Conjugating Enzymes; D044767:Ubiquitin-Protein Ligases; D000071193:Autophagy-Related Protein 7; C104235:ATG3 protein, S cerevisiae
keywords: ATG; Autophagy; Crystallization; Lipidation; Ubiquitin-like proteins