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Identification of Novel Interactions in HIV-1 Capsid Protein Assembly by High-resolution Mass Spectrometry
Abstract
The pleomorphic nature of the immature and mature HIV-1 virions has made it difficult to characterize intersubunit interactions using traditional approaches. While the structures of isolated domains are known, the challenge is to identify intersubunit interactions and thereby pack these domains into supramolecular structures. Using high-resolution mass spectrometry, we have measured the amide hydrogen exchange protection factors for the soluble capsid protein (CA) and CA assembled in vitro. Comparison of the protection factors as well as chemical crosslinking experiments has led to a map of the subunit/subunit interfaces in the assembled tubes. This analysis provides direct biochemical evidence for the homotypic N domain and C domain interactions proposed from cryo-electron microscopy image reconstruction of CA tubes. Most significantly, we have identified a previously unrecognized intersubunit N domain-C domain interaction. The detection of this interaction reconciles previously discrepant biophysical and genetic data.
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“An interface is also present between each CA-CTD and the CA-NTD of the neighbouring CA molecule in the hexameric ring”
From Retrovirus maturation-an extraordinary structural transformation (2016) · cites this paper as [55]
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Metadata
- Authors
- Lanman, J., Lam, T. T., Barnes, S., Sakalian, M., Emmett, M. R., Marshall, A. G., & Prevelige, P. E.
- Affiliations
- University of Alabama at Birmingham (US)
- Florida State University (US)
- University of Oklahoma Health Sciences Center (US)
- Corresponding author
- Prevelige, Peter E.
- Venue
- Journal of Molecular Biology
- Publisher
- Elsevier BV
- Published
- 2003-01
- Volume
- 325
- Issue
- 4
- Pages
- 759-772
- ISSN
- 0022-2836 (ISSN-L)
- OpenAlex
- W1968718594
- Cited by
- 206
- References
- 56
- Citations by year
- 2026: 1 · 2023: 1 · 2022: 3 · 2021: 2 · 2020: 3 · 2019: 3 · 2018: 1 · 2017: 3 · 2016: 2 · 2015: 6 · 2014: 7 · 2013: 14 · 2012: 8
- FWCI
- 5.06
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Journal of Molecular Biology (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Science Foundation
- National High Magnetic Field Laboratory
- National Institutes of Health (5r01gm031683-17)
- National Cancer Institute (T32 CA 09467, T32CA009467)
- National Institute of General Medical Sciences (GM 31683)
- National Institute of Allergy and Infectious Diseases (AI 44626, R01AI044626)
- Topics
- Life Sciences › Immunology and Microbiology › Virology › HIV Research and Treatment
- Physical Sciences › Chemistry › Spectroscopy › Mass Spectrometry Techniques and Applications
- Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
- Keywords
- HIV-1 capsid protein, intersubunit interactions, amide hydrogen exchange, N-domain, C-domain, subunit interface, chemical crosslinking, supramolecular structure
- MeSH terms
- Amides, Cross-Linking Reagents, Deuterium, Hydrogen, Models, Molecular, Mass Spectrometry, HIV-1, Mutagenesis, Site-Directed, Protein Structure, Tertiary, Dimerization, Protein Structure, Quaternary, Protein Subunits, Capsid Proteins, Macromolecular Substances
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
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