{"doi":"10.1016/bs.mie.2016.09.055","title":"Production of Human ATG Proteins for Lipidation Assays","abstract":null,"journal":"Methods in Enzymology","year":2017,"id":657746,"datarank":0.5424726451498179,"base_score":2.302585092994046,"endowment":2.302585092994046,"self_citation_contribution":0.3453877639491069,"citation_network_contribution":0.19708488120071088,"self_endowment_contribution":0.3453877639491069,"citer_contribution":0.19708488120071088,"corpus_percentile":null,"corpus_rank":null,"citation_count":9,"citer_count":5,"citers_with_citation_signal":4,"citers_with_endowment":4,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":null,"is_data_producer":false,"deposit_databanks":null,"is_oa":false,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":null,"fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1717024,"name":"Y. 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Lipidation of LC3 and GABARAP regulates numerous facets of the autophagy process, including regulating expansion of the phagophore membrane, recruiting selected cargoes for degradation, and providing an autophagosome membrane-bound platform mediating dynamic interactions with other regulatory proteins. LC3 and GABARAP are families of related ubiquitin-like proteins (UBLs) (referred to here collectively as LC3/GABARAP), and their lipidation involves a divergent UBL conjugation cascade including ATG7, ATG3, and ATG12~ATG5-ATG16L1 acting as E1, E2, and E3 enzymes, respectively. ATG7 initiates LC3/GABARAP conjugation by catalyzing their C-terminal adenylation and conjugation to the catalytic cysteine of ATG3. Ultimately, the ATG12~ATG5-ATG16L1 complex catalyzes LC3/GABARAP ligation to a primary amino group on PE or other acceptor lipids. This chapter describes methods for expressing and purifying human LC3 or GABARAP, ATG7, ATG3, and the ATG12~ATG5-ATG16L1 complex for in vitro studies of LC3/GABARAP lipidation.","is_dataset_classified":null,"base_score":2.302585092994046,"endowment":2.302585092994046,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"28253979","pmcid":"PMC5450658","openalex_id":"https://openalex.org/W2550929917","authors":[],"funders":[{"funder_name":"NIGMS NIH HHS","grant_id":"R37 GM069530","title":null},{"funder_name":"NIGMS NIH HHS","grant_id":"R01 GM077053","title":null},{"funder_name":"National Institutes of Health","grant_id":"5R01GM077053-07","title":"Structures/mechanisms in a noncanonical ubiquitin-like protein transfer cascade"},{"funder_name":"Howard Hughes Medical Institute","grant_id":"","title":null},{"funder_name":"Howard Hughes Medical Institute","grant_id":"","title":null}],"total_grants":5,"fwci":1.5389,"citation_percentile":0.85135737,"influential_citations":0,"citation_trend":[{"year":2019,"count":1},{"year":2020,"count":3},{"year":2021,"count":3},{"year":2024,"count":1},{"year":2026,"count":1}],"oa_status":"green","license":"Elsevier TDM","oa_locations":[{"url":"https://www.ncbi.nlm.nih.gov/pmc/articles/5450658","host_type":"repository"},{"url":"https://www.ncbi.nlm.nih.gov/pmc/articles/5450658","host_type":"repository"},{"url":"https://api.elsevier.com/content/article/PII:S0076687916303287?httpAccept=text/plain","host_type":"publisher"},{"url":"https://api.elsevier.com/content/article/PII:S0076687916303287?httpAccept=text/xml","host_type":"publisher"},{"url":"https://doi.org/10.1016/bs.mie.2016.09.055","host_type":"book series"},{"url":"https://pubmed.ncbi.nlm.nih.gov/28253979","host_type":"repository"},{"url":"https://europepmc.org/articles/pmc5450658?pdf=render","host_type":""},{"url":"https://dx.doi.org/10.1016/bs.mie.2016.09.055","host_type":""}],"fields_of_study":["Autophagy in Disease and Therapy","Cannabis and Cannabinoid Research","Endoplasmic Reticulum Stress and Disease","0301 basic medicine","0303 health sciences","03 medical and health sciences","Autophagy-Related Proteins","Humans","Microtubule-Associated Proteins","Molecular Biology","Protein Engineering"],"mesh_terms":["Autophagy-Related Proteins","Humans","Microtubule-Associated Proteins","Molecular Biology","Protein Engineering"],"keywords":["Lipid-anchored protein","ATG8","ATG12","ATG5","ATG16L1","Autophagy","Autophagosome","Ubiquitin","Cell biology","ULK1","Chemistry","Biochemistry","Biology","Enzyme","Gene","AMPK","Protein purification","Protein expression","Ubiquitin-like Protein","Lipidation","Autophagy-Related Proteins","Humans","Protein Engineering","Microtubule-Associated Proteins","Molecular Biology"],"sdg_mappings":[{"sdg_number":3,"sdg_label":"3. 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