Bioorthogonal click labeling of an amber-free HIV-1 provirus for in-virus single molecule imaging
Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved..
Structural dynamics of human immunodeficiency virus 1 (HIV-1) envelope (Env) glycoprotein mediate cell entry and facilitate immune evasion. Single-molecule FRET using peptides for Env labeling revealed structural dynamics of Env, but peptide use risks potential effects on structural integrity/dynamics. While incorporating noncanonical amino acids (ncAAs) into Env by amber stop-codon suppression, followed by click chemistry, offers a minimally invasive approach, this has proved to be technically challenging for HIV-1. Here, we develope an intact amber-free HIV-1 system that overcomes hurdles of preexisting viral amber codons. We achieved dual-ncAA incorporation into Env on amber-free virions, enabling single-molecule Förster resonance energy transfer (smFRET) studies of click-labeled Env that validated the previous peptide-based labeling approaches by confirming the intrinsic propensity of Env to dynamically sample multiple conformational states. Amber-free click-labeled Env also enabled real-time tracking of single virion internalization and trafficking in cells. Our system thus permits in-virus bioorthogonal labeling of proteins, compatible with studies of virus entry, trafficking, and egress from cells.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2024 |
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| Erschienen: |
2024 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:31 |
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| Enthalten in: |
Cell chemical biology - 31(2024), 3 vom: 21. März, Seite 487-501.e7 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Ao, Yuanyun [VerfasserIn] |
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| Links: |
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| Anmerkungen: |
Date Completed 25.03.2024 Date Revised 28.03.2024 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1016/j.chembiol.2023.12.017 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| 500 | |a published: Print-Electronic | ||
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| 520 | |a Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved. | ||
| 520 | |a Structural dynamics of human immunodeficiency virus 1 (HIV-1) envelope (Env) glycoprotein mediate cell entry and facilitate immune evasion. Single-molecule FRET using peptides for Env labeling revealed structural dynamics of Env, but peptide use risks potential effects on structural integrity/dynamics. While incorporating noncanonical amino acids (ncAAs) into Env by amber stop-codon suppression, followed by click chemistry, offers a minimally invasive approach, this has proved to be technically challenging for HIV-1. Here, we develope an intact amber-free HIV-1 system that overcomes hurdles of preexisting viral amber codons. We achieved dual-ncAA incorporation into Env on amber-free virions, enabling single-molecule Förster resonance energy transfer (smFRET) studies of click-labeled Env that validated the previous peptide-based labeling approaches by confirming the intrinsic propensity of Env to dynamically sample multiple conformational states. Amber-free click-labeled Env also enabled real-time tracking of single virion internalization and trafficking in cells. Our system thus permits in-virus bioorthogonal labeling of proteins, compatible with studies of virus entry, trafficking, and egress from cells | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a HIV-1 | |
| 650 | 4 | |a amber suppression | |
| 650 | 4 | |a amber-free provirus | |
| 650 | 4 | |a bioorthogonal labeling | |
| 650 | 4 | |a click chemistry | |
| 650 | 4 | |a envelope glycoprotein | |
| 650 | 4 | |a genetic code expansion | |
| 650 | 4 | |a single virus tracking | |
| 650 | 4 | |a single-molecule FRET | |
| 650 | 4 | |a single-molecule imaging | |
| 650 | 7 | |a Proteins |2 NLM | |
| 650 | 7 | |a Peptides |2 NLM | |
| 700 | 1 | |a Grover, Jonathan R |e verfasserin |4 aut | |
| 700 | 1 | |a Gifford, Levi |e verfasserin |4 aut | |
| 700 | 1 | |a Han, Yang |e verfasserin |4 aut | |
| 700 | 1 | |a Zhong, Guohua |e verfasserin |4 aut | |
| 700 | 1 | |a Katte, Revansiddha |e verfasserin |4 aut | |
| 700 | 1 | |a Li, Wenwei |e verfasserin |4 aut | |
| 700 | 1 | |a Bhattacharjee, Rajanya |e verfasserin |4 aut | |
| 700 | 1 | |a Zhang, Baoshan |e verfasserin |4 aut | |
| 700 | 1 | |a Sauve, Stephanie |e verfasserin |4 aut | |
| 700 | 1 | |a Qin, Wenyi |e verfasserin |4 aut | |
| 700 | 1 | |a Ghimire, Dibya |e verfasserin |4 aut | |
| 700 | 1 | |a Haque, Md Anzarul |e verfasserin |4 aut | |
| 700 | 1 | |a Arthos, James |e verfasserin |4 aut | |
| 700 | 1 | |a Moradi, Mahmoud |e verfasserin |4 aut | |
| 700 | 1 | |a Mothes, Walther |e verfasserin |4 aut | |
| 700 | 1 | |a Lemke, Edward A |e verfasserin |4 aut | |
| 700 | 1 | |a Kwong, Peter D |e verfasserin |4 aut | |
| 700 | 1 | |a Melikyan, Gregory B |e verfasserin |4 aut | |
| 700 | 1 | |a Lu, Maolin |e verfasserin |4 aut | |
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