دورية أكاديمية
SUB-immunogold-SEM reveals nanoscale distribution of submembranous epitopes.
| العنوان: | SUB-immunogold-SEM reveals nanoscale distribution of submembranous epitopes. |
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| المؤلفون: | Miller KK; Department of Otolaryngology-Head & Neck Surgery, School of Medicine, 240 Pasteur Drive, Stanford University, Stanford, CA, 94305, USA., Wang P; Department of Otolaryngology-Head & Neck Surgery, School of Medicine, 240 Pasteur Drive, Stanford University, Stanford, CA, 94305, USA., Grillet N; Department of Otolaryngology-Head & Neck Surgery, School of Medicine, 240 Pasteur Drive, Stanford University, Stanford, CA, 94305, USA. ngrillet@stanford.edu. |
| المصدر: | Nature communications [Nat Commun] 2024 Sep 10; Vol. 15 (1), pp. 7864. Date of Electronic Publication: 2024 Sep 10. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Nature Pub. Group Country of Publication: England NLM ID: 101528555 Publication Model: Electronic Cited Medium: Internet ISSN: 2041-1723 (Electronic) Linking ISSN: 20411723 NLM ISO Abbreviation: Nat Commun Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: [London] : Nature Pub. Group |
| مواضيع طبية MeSH: | Epitopes*/immunology , Epitopes*/metabolism , Microscopy, Electron, Scanning*/methods , Immunohistochemistry*/methods , Cilia*/metabolism , Cilia*/ultrastructure, Animals ; Humans ; Hair Cells, Auditory/metabolism ; Hair Cells, Auditory/ultrastructure ; Angiotensin-Converting Enzyme 2/metabolism ; SARS-CoV-2/immunology ; SARS-CoV-2/metabolism ; Myosins/metabolism ; Stereocilia/metabolism ; Stereocilia/ultrastructure ; COVID-19/virology ; COVID-19/immunology |
| مستخلص: | Electron microscopy paired with immunogold labeling is the most precise tool for protein localization. However, these methods are either cumbersome, resulting in small sample numbers and restricted quantification, or limited to identifying protein epitopes external to the membrane. Here, we introduce SUB-immunogold-SEM, a scanning electron microscopy technique that detects intracellular protein epitopes proximal to the membrane. We identify four critical sample preparation factors contributing to the method's sensitivity. We validate its efficacy through precise localization and high-powered quantification of cytoskeletal and transmembrane protein distribution. We evaluate the capabilities of SUB-immunogold-SEM on cells with highly differentiated apical surfaces: (i) auditory hair cells, revealing the presence of nanoscale MYO15A-L rings at the tip of stereocilia; and (ii) respiratory multiciliate cells, mapping the distribution of the SARS-CoV-2 receptor ACE2 along the motile cilia. SUB-immunogold-SEM extends the application of SEM-based nanoscale protein localization to the detection of intracellular epitopes on the exposed surfaces of any cell. (© 2024. The Author(s).) |
| التعليقات: | Update of: Res Sq. 2024 Jan 22:rs.3.rs-3876898. doi: 10.21203/rs.3.rs-3876898/v1. (PMID: 38343799) |
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| معلومات مُعتمدة: | R01 AG081608 United States AG NIA NIH HHS; 1R21DC019457-01 U.S. Department of Health & Human Services | NIH | National Institute on Deafness and Other Communication Disorders (NIDCD); R01 DC016409 United States DC NIDCD NIH HHS; 1R01AG081608-01 U.S. Department of Health & Human Services | NIH | National Institute on Aging (U.S. National Institute on Aging); R21 DC019457 United States DC NIDCD NIH HHS; RO1-DC-016409-01A1 U.S. Department of Health & Human Services | NIH | National Institute on Deafness and Other Communication Disorders (NIDCD) |
| المشرفين على المادة: | 0 (Epitopes) EC 3.4.17.23 (Angiotensin-Converting Enzyme 2) EC 3.6.4.1 (Myosins) EC 3.4.17.23 (ACE2 protein, human) |
| تواريخ الأحداث: | Date Created: 20240910 Date Completed: 20240910 Latest Revision: 20240924 |
| رمز التحديث: | 20240924 |
| مُعرف محوري في PubMed: | PMC11387508 |
| DOI: | 10.1038/s41467-024-51849-x |
| PMID: | 39256352 |
| قاعدة البيانات: | MEDLINE |
Find this article in full text from Springer Verlag. 
Full Text Finder | تدمد: | 2041-1723 |
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| DOI: | 10.1038/s41467-024-51849-x |