Cargando…

Targeting the Structural Maturation Pathway of HIV-1 Reverse Transcriptase

Formation of active HIV-1 reverse transcriptase (RT) proceeds via a structural maturation process that involves subdomain rearrangements and formation of an asymmetric p66/p66′ homodimer. These studies were undertaken to evaluate whether the information about this maturation process can be used to i...

Descripción completa

Detalles Bibliográficos
Autores principales: Kirby, Thomas W., Gabel, Scott A., DeRose, Eugene F., Perera, Lalith, Krahn, Juno M., Pedersen, Lars C., London, Robert E.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10669680/
https://www.ncbi.nlm.nih.gov/pubmed/38002285
http://dx.doi.org/10.3390/biom13111603
_version_ 1785139753426878464
author Kirby, Thomas W.
Gabel, Scott A.
DeRose, Eugene F.
Perera, Lalith
Krahn, Juno M.
Pedersen, Lars C.
London, Robert E.
author_facet Kirby, Thomas W.
Gabel, Scott A.
DeRose, Eugene F.
Perera, Lalith
Krahn, Juno M.
Pedersen, Lars C.
London, Robert E.
author_sort Kirby, Thomas W.
collection PubMed
description Formation of active HIV-1 reverse transcriptase (RT) proceeds via a structural maturation process that involves subdomain rearrangements and formation of an asymmetric p66/p66′ homodimer. These studies were undertaken to evaluate whether the information about this maturation process can be used to identify small molecule ligands that retard or interfere with the steps involved. We utilized the isolated polymerase domain, p51, rather than p66, since the initial subdomain rearrangements are largely limited to this domain. Target sites at subdomain interfaces were identified and computational analysis used to obtain an initial set of ligands for screening. Chromatographic evaluations of the p51 homodimer/monomer ratio support the feasibility of this approach. Ligands that bind near the interfaces and a ligand that binds directly to a region of the fingers subdomain involved in subunit interface formation were identified, and the interactions were further characterized by NMR spectroscopy and X-ray crystallography. Although these ligands were found to reduce dimer formation, further efforts will be required to obtain ligands with higher binding affinity. In contrast with previous ligand identification studies performed on the RT heterodimer, subunit interface surfaces are solvent-accessible in the p51 and p66 monomers, making these constructs preferable for identification of ligands that directly interfere with dimerization.
format Online
Article
Text
id pubmed-10669680
institution National Center for Biotechnology Information
language English
publishDate 2023
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-106696802023-11-01 Targeting the Structural Maturation Pathway of HIV-1 Reverse Transcriptase Kirby, Thomas W. Gabel, Scott A. DeRose, Eugene F. Perera, Lalith Krahn, Juno M. Pedersen, Lars C. London, Robert E. Biomolecules Article Formation of active HIV-1 reverse transcriptase (RT) proceeds via a structural maturation process that involves subdomain rearrangements and formation of an asymmetric p66/p66′ homodimer. These studies were undertaken to evaluate whether the information about this maturation process can be used to identify small molecule ligands that retard or interfere with the steps involved. We utilized the isolated polymerase domain, p51, rather than p66, since the initial subdomain rearrangements are largely limited to this domain. Target sites at subdomain interfaces were identified and computational analysis used to obtain an initial set of ligands for screening. Chromatographic evaluations of the p51 homodimer/monomer ratio support the feasibility of this approach. Ligands that bind near the interfaces and a ligand that binds directly to a region of the fingers subdomain involved in subunit interface formation were identified, and the interactions were further characterized by NMR spectroscopy and X-ray crystallography. Although these ligands were found to reduce dimer formation, further efforts will be required to obtain ligands with higher binding affinity. In contrast with previous ligand identification studies performed on the RT heterodimer, subunit interface surfaces are solvent-accessible in the p51 and p66 monomers, making these constructs preferable for identification of ligands that directly interfere with dimerization. MDPI 2023-11-01 /pmc/articles/PMC10669680/ /pubmed/38002285 http://dx.doi.org/10.3390/biom13111603 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Kirby, Thomas W.
Gabel, Scott A.
DeRose, Eugene F.
Perera, Lalith
Krahn, Juno M.
Pedersen, Lars C.
London, Robert E.
Targeting the Structural Maturation Pathway of HIV-1 Reverse Transcriptase
title Targeting the Structural Maturation Pathway of HIV-1 Reverse Transcriptase
title_full Targeting the Structural Maturation Pathway of HIV-1 Reverse Transcriptase
title_fullStr Targeting the Structural Maturation Pathway of HIV-1 Reverse Transcriptase
title_full_unstemmed Targeting the Structural Maturation Pathway of HIV-1 Reverse Transcriptase
title_short Targeting the Structural Maturation Pathway of HIV-1 Reverse Transcriptase
title_sort targeting the structural maturation pathway of hiv-1 reverse transcriptase
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10669680/
https://www.ncbi.nlm.nih.gov/pubmed/38002285
http://dx.doi.org/10.3390/biom13111603
work_keys_str_mv AT kirbythomasw targetingthestructuralmaturationpathwayofhiv1reversetranscriptase
AT gabelscotta targetingthestructuralmaturationpathwayofhiv1reversetranscriptase
AT deroseeugenef targetingthestructuralmaturationpathwayofhiv1reversetranscriptase
AT pereralalith targetingthestructuralmaturationpathwayofhiv1reversetranscriptase
AT krahnjunom targetingthestructuralmaturationpathwayofhiv1reversetranscriptase
AT pedersenlarsc targetingthestructuralmaturationpathwayofhiv1reversetranscriptase
AT londonroberte targetingthestructuralmaturationpathwayofhiv1reversetranscriptase