Influence of the heme distal pocket on nitrite binding orientation and reactivity in Sperm Whale myoglobin

Tse, Wilford, Whitmore, Nathan, Cheesman, Myles R. and Watmough, Nicholas J. ORCID: https://orcid.org/0000-0001-5901-6750 (2021) Influence of the heme distal pocket on nitrite binding orientation and reactivity in Sperm Whale myoglobin. Biochemical Journal, 478 (4). pp. 927-942. ISSN 0264-6021

[thumbnail of Published_Version]
Preview
PDF (Published_Version) - Published Version
Available under License Creative Commons Attribution.

Download (3MB) | Preview

Abstract

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.

Item Type: Article
Faculty \ School: Faculty of Science > School of Chemistry
Faculty of Science > School of Biological Sciences
Related URLs:
Depositing User: LivePure Connector
Date Deposited: 05 Mar 2021 00:44
Last Modified: 23 Oct 2022 02:16
URI: https://ueaeprints.uea.ac.uk/id/eprint/79380
DOI: 10.1042/BCJ20200596

Actions (login required)

View Item View Item