ArticleProceedings of the National Academy of Sciences of the United States of America2021
An in vitro vesicle formation assay reveals cargo clients and factors that mediate vesicular trafficking.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 30 papers.
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Who cites it
30 citing papers in PubMed, 45 citations in OpenAlex.
- Zinc-mediated structural and functional regulation of ERp44 and ERGIC-53 in protein quality control.Journal of biochemistry · 2026Review
- Synphilin-1 mitigates autophagy dysfusnction, modulates ubiquitinated protein aggregation, and promotes cell survival during proteotoxic stress.bioRxiv : the preprint server for biology · 2026Article
- Tissue-selective COPII modulator SEC16B aggravates cardiovascular disease by promoting lipid export.The EMBO journal · 2026Article
- Article
- Mechanisms of COPII coat assembly and cargo recognition in the secretory pathway.Nature reviews. Molecular cell biology · 2025Review
- Public Cohort Analysis Identifies Thyroglobulin Variants as Hypothyroidism Risk Factors.bioRxiv : the preprint server for biology · 2025Article
- Uncovering cargo clients and accessory factors of AP-1 and AP-4 through vesicle proteomics.Proceedings of the National Academy of Sciences of the United States of America · 2025Article
- GAS2L1 facilitates Golgi-derived microtubule growth and Golgi organization.Molecular biology of the cell · 2025Article
- ST6GalNAc-I regulates tumor cell sialylation via NECTIN2/MUC5AC-mediated immunosuppression and angiogenesis in non-small cell lung cancer.The Journal of clinical investigation · 2025Article
- Characterization of extracellular vesicles released fromPhilosophical transactions of the Royal Society of London. Series B, Biological sciences · 2025Article
- ER export via SURF4 uses diverse mechanisms of both client and coat engagement.The Journal of cell biology · 2025Article
- Endoplasmic reticulum exit sites are segregated for secretion based on cargo size.Developmental cell · 2024Article
- The ufmylation cascade controls COPII recruitment, anterograde transport, and sorting of nascent GPCRs at ER.Science advances · 2024Article
- Structure of full-length ERGIC-53 in complex with MCFD2 for cargo transport.Nature communications · 2024Article
- In Vitro Reconstitution of Plant COPII Vesicles.Methods in molecular biology (Clifton, N.J.) · 2024Article
- Proteomic Analysis of Secreted Vesicle Proteins from the Plant Cells.Methods in molecular biology (Clifton, N.J.) · 2024Article
- Nlp-dependent ER-to-Golgi transport.International journal of biological sciences · 2024Article
- TMED10 mediates the trafficking of insulin-like growth factor 2 along the secretory pathway for myoblast differentiation.Proceedings of the National Academy of Sciences of the United States of America · 2023Article
- Identification of LMAN1- and SURF4-Dependent Secretory Cargoes.Journal of proteome research · 2023Article
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Authors and funding
14 authors at 4 institutions in 3 countries.
Funding
Abstract
The fidelity of protein transport in the secretory pathway relies on the accurate sorting of proteins to their correct destinations. To deepen our understanding of the underlying molecular mechanisms, it is important to develop a robust approach to systematically reveal cargo proteins that depend on specific sorting machinery to be enriched into transport vesicles. Here, we used an in vitro assay that reconstitutes packaging of human cargo proteins into vesicles to quantify cargo capture. Quantitative mass spectrometry (MS) analyses of the isolated vesicles revealed cytosolic proteins that are associated with vesicle membranes in a GTP-dependent manner. We found that two of them, FAM84B (also known as LRAT domain containing 2 or LRATD2) and PRRC1, contain proline-rich domains and regulate anterograde trafficking. Further analyses revealed that PRRC1 is recruited to endoplasmic reticulum (ER) exit sites, interacts with the inner COPII coat, and its absence increases membrane association of COPII. In addition, we uncovered cargo proteins that depend on GTP hydrolysis to be captured into vesicles. Comparing control cells with cells depleted of the cargo receptors, SURF4 or ERGIC53, we revealed specific clients of each of these two export adaptors. Our results indicate that the vesicle formation assay in combination with quantitative MS analysis is a robust and powerful tool to uncover novel factors that mediate vesicular trafficking and to uncover cargo clients of specific cellular factors.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.