ArticleNature protocols2015
Extracellular protease digestion to evaluate membrane protein cell surface localization.
Article in Nature protocols, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 23 papers.
What it found
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.
Who cites it
23 citing papers in PubMed.
- Exosomes in Amyloid Propagation-Roles in Neurodegeneration.Molecular neurobiology · 2026Review
- Dynamic translocation of Inside-Out proteins to the cell surface underlies cellular adaptation to cancer-induced stress.Proceedings of the National Academy of Sciences of the United States of America · 2026Article
- Cell Surface Shaving-Based Proteomic Profiling of the Surfaceome in Pathogenic Microorganisms.International journal of molecular sciences · 2026Review
- Divergence in surface protein exposure between reference and clinical-derivedActa biochimica Polonica · 2026Article
- The biochemical function of bivalent aptamer assemblies against B cell markers CD19 and CD20.Molecular therapy. Nucleic acids · 2025Article
- Ampicillin treatment in persister cell studies may cause non-physiological artifacts.Microbial cell (Graz, Austria) · 2025Article
- MoAti1 mediates mitophagy by facilitating recruitment of MoAtg8 to promote invasive growth in Magnaporthe oryzae.Molecular plant pathology · 2024Article
- Structural analysis of platelet fragments and extracellular vesicles produced by apheresis platelets during storage.Blood advances · 2024Article
- Programming Cell-Derived Vesicles with Enhanced Immunomodulatory Properties.Advanced healthcare materials · 2023Article
- Characterization of Treponema denticola Major Surface Protein (Msp) by Deletion Analysis and Advanced Molecular Modeling.Journal of bacteriology · 2022Article
- Keratin 8 is a scaffolding and regulatory protein of ERAD complexes.Cellular and molecular life sciences : CMLS · 2022Article
- Two trehalase isoforms, produced from a single transcript, regulate drought stress tolerance in Arabidopsis thaliana.Plant molecular biology · 2022Article
- Colicin E1 opens its hinge to plug TolC.eLife · 2022Article
- Lipid bilayer induces contraction of the denatured state ensemble of a helical-bundle membrane protein.Proceedings of the National Academy of Sciences of the United States of America · 2022Article
- Engineering Tropism ofACS synthetic biology · 2021Article
- Review
- Current Status and Perspectives of Protease Inhibitors and Their Combination with Nanosized Drug Delivery Systems for Targeted Cancer Therapy.Drug design, development and therapy · 2021Review
- Dead cells release a 'necrosignal' that activates antibiotic survival pathways in bacterial swarms.Nature communications · 2020Article
- Phenotypic traits of mesenchymal stem cell sheets fabricated by temperature-responsive cell culture plate: structural characteristics of MSC sheets.Stem cell research & therapy · 2019Article
- Bacterial killing by complement requires membrane attack complex formation via surface-bound C5 convertases.The EMBO journal · 2019Article
Corrections and comments
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Authors and funding
2 authors.
Funding
Abstract
Membrane proteins have crucial roles in signaling and as anchors for cell surface display. Proper secretion of a membrane protein can be evaluated by its susceptibility to digestion by an extracellular protease, but this requires a crucial control to confirm membrane integrity during digestion. This protocol describes how to use this approach to determine how efficiently a protein is secreted to the outer surface of Gram-negative bacteria. Its success relies upon careful selection of an appropriate intracellular reporter protein that will remain undigested if the membrane barrier remains intact, but that is rapidly digested when cells are lysed before evaluation. Reporter proteins that are resistant to proteases (e.g., maltose-binding protein) do not return accurate results; in contrast, proteins that are more readily digested (e.g., SurA) serve as more sensitive reporters of membrane integrity, yielding more accurate measurements of membrane protein localization. Similar considerations apply when evaluating membrane protein localization in other contexts, including eukaryotic cells and organelle membranes. Evaluating membrane protein localization using this approach requires only standard biochemistry laboratory equipment for cell lysis, gel electrophoresis and western blotting. After expression of the protein of interest, this procedure can be completed in 4 h.
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Registered trials
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.