Influence of the heme distal pocket on nitrite binding orientation and reactivity in Sperm Whale myoglobin
Nitrite binding to recombinant wild-type Sperm Whale myoglobin (SWMb) was studied using a combination of spectroscopic methods including room-temperature magnetic circular dichroism. These revealed that the reactive species is free nitrous acid and the product of the reaction contains a nitrite ion...
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Portland Press Ltd.
2021
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crportlandpress:10.1042/bcj20200596 2024-06-23T07:56:58+00:00 Influence of the heme distal pocket on nitrite binding orientation and reactivity in Sperm Whale myoglobin Tse, Wilford Whitmore, Nathan Cheesman, Myles R. Watmough, Nicholas J. 2021 http://dx.doi.org/10.1042/bcj20200596 https://portlandpress.com/biochemj/article-pdf/478/4/927/904911/bcj-2020-0596.pdf en eng Portland Press Ltd. https://creativecommons.org/licenses/by/4.0/ Biochemical Journal volume 478, issue 4, page 927-942 ISSN 0264-6021 1470-8728 journal-article 2021 crportlandpress https://doi.org/10.1042/bcj20200596 2024-05-30T08:21:55Z Nitrite binding to recombinant wild-type Sperm Whale myoglobin (SWMb) was studied using a combination of spectroscopic methods including room-temperature magnetic circular dichroism. These revealed that the reactive species is free nitrous acid and the product of the reaction contains a nitrite ion bound to the ferric heme iron in the nitrito- (O-bound) orientation. This exists in a thermal equilibrium with a low-spin ground state and a high-spin excited state and is spectroscopically distinct from the purely low-spin nitro- (N-bound) species observed in the H64V SWMb variant. Substitution of the proximal heme ligand, histidine-93, with lysine yields a novel form of myoglobin (H93K) with enhanced reactivity towards nitrite. The nitrito-mode of binding to the ferric heme iron is retained in the H93K variant again as a thermal equilibrium of spin-states. This proximal substitution influences the heme distal pocket causing the pKa of the alkaline transition to be lowered relative to wild-type SWMb. This change in the environment of the distal pocket coupled with nitrito-binding is the most likely explanation for the 8-fold increase in the rate of nitrite reduction by H93K relative to WT SWMb. Article in Journal/Newspaper Sperm whale Portland Press Biochemical Journal 478 4 927 942 |
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Open Polar |
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Portland Press |
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English |
description |
Nitrite binding to recombinant wild-type Sperm Whale myoglobin (SWMb) was studied using a combination of spectroscopic methods including room-temperature magnetic circular dichroism. These revealed that the reactive species is free nitrous acid and the product of the reaction contains a nitrite ion bound to the ferric heme iron in the nitrito- (O-bound) orientation. This exists in a thermal equilibrium with a low-spin ground state and a high-spin excited state and is spectroscopically distinct from the purely low-spin nitro- (N-bound) species observed in the H64V SWMb variant. Substitution of the proximal heme ligand, histidine-93, with lysine yields a novel form of myoglobin (H93K) with enhanced reactivity towards nitrite. The nitrito-mode of binding to the ferric heme iron is retained in the H93K variant again as a thermal equilibrium of spin-states. This proximal substitution influences the heme distal pocket causing the pKa of the alkaline transition to be lowered relative to wild-type SWMb. This change in the environment of the distal pocket coupled with nitrito-binding is the most likely explanation for the 8-fold increase in the rate of nitrite reduction by H93K relative to WT SWMb. |
format |
Article in Journal/Newspaper |
author |
Tse, Wilford Whitmore, Nathan Cheesman, Myles R. Watmough, Nicholas J. |
spellingShingle |
Tse, Wilford Whitmore, Nathan Cheesman, Myles R. Watmough, Nicholas J. Influence of the heme distal pocket on nitrite binding orientation and reactivity in Sperm Whale myoglobin |
author_facet |
Tse, Wilford Whitmore, Nathan Cheesman, Myles R. Watmough, Nicholas J. |
author_sort |
Tse, Wilford |
title |
Influence of the heme distal pocket on nitrite binding orientation and reactivity in Sperm Whale myoglobin |
title_short |
Influence of the heme distal pocket on nitrite binding orientation and reactivity in Sperm Whale myoglobin |
title_full |
Influence of the heme distal pocket on nitrite binding orientation and reactivity in Sperm Whale myoglobin |
title_fullStr |
Influence of the heme distal pocket on nitrite binding orientation and reactivity in Sperm Whale myoglobin |
title_full_unstemmed |
Influence of the heme distal pocket on nitrite binding orientation and reactivity in Sperm Whale myoglobin |
title_sort |
influence of the heme distal pocket on nitrite binding orientation and reactivity in sperm whale myoglobin |
publisher |
Portland Press Ltd. |
publishDate |
2021 |
url |
http://dx.doi.org/10.1042/bcj20200596 https://portlandpress.com/biochemj/article-pdf/478/4/927/904911/bcj-2020-0596.pdf |
genre |
Sperm whale |
genre_facet |
Sperm whale |
op_source |
Biochemical Journal volume 478, issue 4, page 927-942 ISSN 0264-6021 1470-8728 |
op_rights |
https://creativecommons.org/licenses/by/4.0/ |
op_doi |
https://doi.org/10.1042/bcj20200596 |
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Biochemical Journal |
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478 |
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4 |
container_start_page |
927 |
op_container_end_page |
942 |
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1802650382103478272 |