{"doi":"10.1083/jcb.147.7.1457","title":"ER/Golgi Intermediates Acquire Golgi Enzymes by Brefeldin a–Sensitive Retrograde Transport in Vitro","abstract":"<jats:p>Secretory proteins exit the ER in transport vesicles that fuse to form vesicular tubular clusters (VTCs) which move along microtubule tracks to the Golgi apparatus. Using the well-characterized in vitro approach to study the properties of Golgi membranes, we determined whether the Golgi enzyme NAGT I is transported to ER/Golgi intermediates. Secretory cargo was arrested at distinct steps of the secretory pathway of a glycosylation mutant cell line, and in vitro complementation of the glycosylation defect was determined. Complementation yield increased after ER exit of secretory cargo and was optimal when transport was blocked at an ER/Golgi intermediate step. The rapid drop of the complementation yield as secretory cargo progresses into the stack suggests that Golgi enzymes are preferentially targeted to ER/Golgi intermediates and not to membranes of the Golgi stack. Two mechanisms for in vitro complementation could be distinguished due to their different sensitivities to brefeldin A (BFA). Transport occurred either by direct fusion of preexisting transport intermediates with ER/Golgi intermediates, or it occurred as a BFA-sensitive and most likely COP I–mediated step. Direct fusion of ER/Golgi intermediates with cisternal membranes of the Golgi stack was not observed under these conditions.</jats:p>","journal":"The Journal of Cell Biology","year":1999,"id":31137,"datarank":1.8492638293557468,"base_score":3.6635616461296463,"endowment":3.6635616461296463,"self_citation_contribution":0.5495342469194471,"citation_network_contribution":1.2997295824362998,"self_endowment_contribution":0.5495342469194471,"citer_contribution":1.2997295824362998,"corpus_percentile":null,"corpus_rank":null,"citation_count":38,"citer_count":34,"citers_with_citation_signal":32,"citers_with_endowment":32,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":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":167538,"name":"Harold D. Love","orcid":null,"position":1,"is_corresponding":false},{"id":167539,"name":"Jennifer N. Gushue","orcid":null,"position":2,"is_corresponding":false},{"id":167540,"name":"John J.M. Bergeron","orcid":null,"position":3,"is_corresponding":false},{"id":167541,"name":"Joachim Ostermann","orcid":null,"position":4,"is_corresponding":false},{"id":167537,"name":"Chung-Chih Lin","orcid":null,"position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"base_score":3.6635616461296463,"endowment":3.6635616461296463,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"10613904","pmcid":"PMC2174244","openalex_id":"https://openalex.org/W2080472611","authors":[],"funders":[],"total_grants":0,"fwci":1.5569,"citation_percentile":0.82158337,"influential_citations":2,"citation_trend":[{"year":2012,"count":1},{"year":2014,"count":1},{"year":2015,"count":2},{"year":2017,"count":1},{"year":2018,"count":1},{"year":2021,"count":1},{"year":2022,"count":1},{"year":2023,"count":1}],"oa_status":"bronze","license":null,"oa_locations":[{"url":"http://jcb.rupress.org/content/147/7/1457.full.pdf","host_type":"journal"},{"url":"http://jcb.rupress.org/content/147/7/1457.full.pdf","host_type":"BRONZE"},{"url":"http://jcb.rupress.org/content/147/7/1457.full.pdf","host_type":"publisher"},{"url":"https://rupress.org/jcb/article-pdf/147/7/1457/1857162/9907098.pdf","host_type":"publisher"},{"url":"https://doi.org/10.1083/jcb.147.7.1457","host_type":"journal"},{"url":"https://pubmed.ncbi.nlm.nih.gov/10613904","host_type":"repository"},{"url":"http://citeseerx.ist.psu.edu/viewdoc/summary?doi=10.1.1.273.7413","host_type":""},{"url":"https://www.ncbi.nlm.nih.gov/pmc/articles/2174244","host_type":"repository"},{"url":"https://europepmc.org/articles/PMC2174244","host_type":"Europe_PMC"},{"url":"https://europepmc.org/articles/PMC2174244?pdf=render","host_type":"Europe_PMC"}],"fields_of_study":["Cellular transport and secretion","Lipid Membrane Structure and Behavior","Lysosomal Storage Disorders Research","Biology","Medicine","Animals","Antibodies","Biological Transport","Brefeldin A","CHO Cells","Centrifugation, Density Gradient","Coatomer Protein","Cricetinae","Endoplasmic Reticulum","Genetic Complementation Test","Glycosylation","Golgi Apparatus","Intracellular Membranes","Membrane Glycoproteins","Membrane Proteins","Microscopy, Immunoelectron","Temperature","Time Factors","Vesicular stomatitis Indiana virus","Viral Envelope Proteins"],"mesh_terms":["Animals","Antibodies","Biological Transport","Centrifugation, Density Gradient","Endoplasmic Reticulum","Genetic Complementation Test","Glycosylation","Golgi Apparatus","Cricetinae","Intracellular Membranes","Membrane Glycoproteins","Membrane Proteins","Temperature","Time Factors","Vesicular stomatitis Indiana virus","Viral Envelope Proteins","Microscopy, Immunoelectron","CHO Cells","Brefeldin A","Coatomer Protein"],"keywords":["Golgi apparatus","Brefeldin A","Secretory pathway","Cell biology","Biology","Complementation","Vesicle","Endosome","Exocytosis","Golgi membrane","Secretion","Endoplasmic reticulum","Biochemistry","Membrane","Mutant","Intracellular"],"sdg_mappings":[{"sdg_number":0,"sdg_label":"Life below water"}],"linked_datasets":[],"clinical_trials":[],"software_tools":[],"database_accessions":[],"source":"live","citation_network_status":"fetched"},"created_at":"2026-06-09T06:34:17.271391Z","pmid":null,"pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}