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Corrole–protein interactions in H-NOX and HasA

Replacing the native porphyrin cofactor in haem proteins has led to the development of novel designer proteins for a variety of applications. In most cases, haem analogues bind in a way that is comparable to the iron porphyrin, but this is not necessarily the case for complexes bearing non-exchangea...

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Autores principales: Lemon, Christopher M., Nissley, Amos J., Latorraca, Naomi R., Wittenborn, Elizabeth C., Marletta, Michael A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: RSC 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9092467/
https://www.ncbi.nlm.nih.gov/pubmed/35656484
http://dx.doi.org/10.1039/d2cb00004k
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author Lemon, Christopher M.
Nissley, Amos J.
Latorraca, Naomi R.
Wittenborn, Elizabeth C.
Marletta, Michael A.
author_facet Lemon, Christopher M.
Nissley, Amos J.
Latorraca, Naomi R.
Wittenborn, Elizabeth C.
Marletta, Michael A.
author_sort Lemon, Christopher M.
collection PubMed
description Replacing the native porphyrin cofactor in haem proteins has led to the development of novel designer proteins for a variety of applications. In most cases, haem analogues bind in a way that is comparable to the iron porphyrin, but this is not necessarily the case for complexes bearing non-exchangeable ligands. This study probes how a P[double bond, length as m-dash]O corrole binds to functionally disparate hemoproteins: a haem-dependent oxygen sensor (H-NOX) and a haem-scavenging protein (HasA). The results demonstrate that the protein–cofactor interactions are distinct from the native, haem-bound holoprotein. In H-NOX, the P[double bond, length as m-dash]O unit primarily hydrogen bonds with the haem-ligating histidine (H102), rather than the hydrogen-bonding network that stabilises the Fe(ii)–O(2) complex in the native protein. In the absence of H102, the protein is still able to bind the corrole, albeit at reduced levels. Molecular dynamics simulations were utilised to determine the flexibility of apo H-NOX and revealed the coupled motion of key residues necessary for corrole binding. In the case of HasA, the P[double bond, length as m-dash]O unit does not primarily interact with either the haem-ligating histidine (H32) or tyrosine (Y75). Instead, histidine 83, the hydrogen-bonding partner for Y75, is critical for P[double bond, length as m-dash]O corrole binding. The conformation of HasA is interrogated by site-specifically labelling the protein and exploiting Förster resonance energy transfer (FRET) to determine the dye–cofactor distance. HasA reconstituted with the P[double bond, length as m-dash]O corrole exhibits an extended, apo-like conformation. Together, these results demonstrate that non-natural cofactors can bind to proteins in unexpected ways and highlight the need to uncover these interactions for the further development of designer haem proteins.
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spelling pubmed-90924672022-06-01 Corrole–protein interactions in H-NOX and HasA Lemon, Christopher M. Nissley, Amos J. Latorraca, Naomi R. Wittenborn, Elizabeth C. Marletta, Michael A. RSC Chem Biol Chemistry Replacing the native porphyrin cofactor in haem proteins has led to the development of novel designer proteins for a variety of applications. In most cases, haem analogues bind in a way that is comparable to the iron porphyrin, but this is not necessarily the case for complexes bearing non-exchangeable ligands. This study probes how a P[double bond, length as m-dash]O corrole binds to functionally disparate hemoproteins: a haem-dependent oxygen sensor (H-NOX) and a haem-scavenging protein (HasA). The results demonstrate that the protein–cofactor interactions are distinct from the native, haem-bound holoprotein. In H-NOX, the P[double bond, length as m-dash]O unit primarily hydrogen bonds with the haem-ligating histidine (H102), rather than the hydrogen-bonding network that stabilises the Fe(ii)–O(2) complex in the native protein. In the absence of H102, the protein is still able to bind the corrole, albeit at reduced levels. Molecular dynamics simulations were utilised to determine the flexibility of apo H-NOX and revealed the coupled motion of key residues necessary for corrole binding. In the case of HasA, the P[double bond, length as m-dash]O unit does not primarily interact with either the haem-ligating histidine (H32) or tyrosine (Y75). Instead, histidine 83, the hydrogen-bonding partner for Y75, is critical for P[double bond, length as m-dash]O corrole binding. The conformation of HasA is interrogated by site-specifically labelling the protein and exploiting Förster resonance energy transfer (FRET) to determine the dye–cofactor distance. HasA reconstituted with the P[double bond, length as m-dash]O corrole exhibits an extended, apo-like conformation. Together, these results demonstrate that non-natural cofactors can bind to proteins in unexpected ways and highlight the need to uncover these interactions for the further development of designer haem proteins. RSC 2022-03-21 /pmc/articles/PMC9092467/ /pubmed/35656484 http://dx.doi.org/10.1039/d2cb00004k Text en This journal is © The Royal Society of Chemistry https://creativecommons.org/licenses/by-nc/3.0/
spellingShingle Chemistry
Lemon, Christopher M.
Nissley, Amos J.
Latorraca, Naomi R.
Wittenborn, Elizabeth C.
Marletta, Michael A.
Corrole–protein interactions in H-NOX and HasA
title Corrole–protein interactions in H-NOX and HasA
title_full Corrole–protein interactions in H-NOX and HasA
title_fullStr Corrole–protein interactions in H-NOX and HasA
title_full_unstemmed Corrole–protein interactions in H-NOX and HasA
title_short Corrole–protein interactions in H-NOX and HasA
title_sort corrole–protein interactions in h-nox and hasa
topic Chemistry
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9092467/
https://www.ncbi.nlm.nih.gov/pubmed/35656484
http://dx.doi.org/10.1039/d2cb00004k
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