Mechanistic insight into the nitrosylation of the [4Fe−4S] cluster of WhiB-like proteins

Jason C. Crack; Laura J. Smith; Melanie R. Stapleton; Jamie Peck; Nicholas J. Watmough; Mark J. Buttner; Roger S. Buxton; Jeffrey Green; Vasily S. Oganesyan; Andrew J. Thomson; Nick E. Le Brun

doi:10.1021/ja109581t

Mechanistic insight into the nitrosylation of the [4Fe−4S] cluster of WhiB-like proteins

Jason C. Crack, Laura J. Smith, Melanie R. Stapleton, Jamie Peck, Nicholas J. Watmough, Mark J. Buttner, Roger S. Buxton, Jeffrey Green, Vasily S. Oganesyan, Andrew J. Thomson, Nick E. Le Brun

Research output: Contribution to journal › Article › peer-review

123 Citations (Scopus)

Abstract

The reactivity of protein bound iron-sulfur clusters with nitric oxide (NO) is well documented, but little is known about the actual mechanism of cluster nitrosylation. Here, we report studies of members of the Wbl family of [4Fe-4S] containing proteins, which play key roles in regulating developmental processes in actinomycetes, including Streptomyces and Mycobacteria, and have been shown to be NO responsive. Streptomyces coelicolor WhiD and Mycobacterium tuberculosis WhiB1 react extremely rapidly with NO in a multiphasic reaction involving, remarkably, 8 NO molecules per [4Fe-4S] cluster. The reaction is 10⁴-fold faster than that observed with O₂ and is by far the most rapid iron-sulfur cluster nitrosylation reaction reported to date. An overall stoichiometry of [Fe₄S₄(Cys)₄]^2- + 8NO ? 2[Fe^I₂(NO)₄(Cys)₂]⁰ + S^2- + 3S⁰ has been established by determination of the sulfur products and their oxidation states. Kinetic analysis leads to a four-step mechanism that accounts for the observed NO dependence. DFT calculations suggest the possibility that the nitrosylation product is a novel cluster [Fe^I₄(NO)₈(Cys)₄]⁰ derived by dimerization of a pair of Roussin’s red ester (RRE) complexes.

Original language	English
Pages (from-to)	1112-1121
Number of pages	10
Journal	Journal of the American Chemical Society
Volume	133
Issue number	4
DOIs	https://doi.org/10.1021/ja109581t
Publication status	Published - 2011

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.1021/ja109581t

Cite this

@article{137d0dd6ecea4fd5bc3658e554b4a679,

title = "Mechanistic insight into the nitrosylation of the [4Fe−4S] cluster of WhiB-like proteins",

abstract = "The reactivity of protein bound iron-sulfur clusters with nitric oxide (NO) is well documented, but little is known about the actual mechanism of cluster nitrosylation. Here, we report studies of members of the Wbl family of [4Fe-4S] containing proteins, which play key roles in regulating developmental processes in actinomycetes, including Streptomyces and Mycobacteria, and have been shown to be NO responsive. Streptomyces coelicolor WhiD and Mycobacterium tuberculosis WhiB1 react extremely rapidly with NO in a multiphasic reaction involving, remarkably, 8 NO molecules per [4Fe-4S] cluster. The reaction is 104-fold faster than that observed with O2 and is by far the most rapid iron-sulfur cluster nitrosylation reaction reported to date. An overall stoichiometry of [Fe4S4(Cys)4]2- + 8NO ? 2[FeI2(NO)4(Cys)2]0 + S2- + 3S0 has been established by determination of the sulfur products and their oxidation states. Kinetic analysis leads to a four-step mechanism that accounts for the observed NO dependence. DFT calculations suggest the possibility that the nitrosylation product is a novel cluster [FeI4(NO)8(Cys)4]0 derived by dimerization of a pair of Roussin{\textquoteright}s red ester (RRE) complexes.",

author = "Crack, {Jason C.} and Smith, {Laura J.} and Stapleton, {Melanie R.} and Jamie Peck and Watmough, {Nicholas J.} and Buttner, {Mark J.} and Buxton, {Roger S.} and Jeffrey Green and Oganesyan, {Vasily S.} and Thomson, {Andrew J.} and {Le Brun}, {Nick E.}",

year = "2011",

doi = "10.1021/ja109581t",

language = "English",

volume = "133",

pages = "1112--1121",

journal = "Journal of the American Chemical Society",

issn = "0002-7863",

publisher = "American Chemical Society",

number = "4",

}

TY - JOUR

T1 - Mechanistic insight into the nitrosylation of the [4Fe−4S] cluster of WhiB-like proteins

AU - Crack, Jason C.

AU - Smith, Laura J.

AU - Stapleton, Melanie R.

AU - Peck, Jamie

AU - Watmough, Nicholas J.

AU - Buttner, Mark J.

AU - Buxton, Roger S.

AU - Green, Jeffrey

AU - Oganesyan, Vasily S.

AU - Thomson, Andrew J.

AU - Le Brun, Nick E.

PY - 2011

Y1 - 2011

N2 - The reactivity of protein bound iron-sulfur clusters with nitric oxide (NO) is well documented, but little is known about the actual mechanism of cluster nitrosylation. Here, we report studies of members of the Wbl family of [4Fe-4S] containing proteins, which play key roles in regulating developmental processes in actinomycetes, including Streptomyces and Mycobacteria, and have been shown to be NO responsive. Streptomyces coelicolor WhiD and Mycobacterium tuberculosis WhiB1 react extremely rapidly with NO in a multiphasic reaction involving, remarkably, 8 NO molecules per [4Fe-4S] cluster. The reaction is 104-fold faster than that observed with O2 and is by far the most rapid iron-sulfur cluster nitrosylation reaction reported to date. An overall stoichiometry of [Fe4S4(Cys)4]2- + 8NO ? 2[FeI2(NO)4(Cys)2]0 + S2- + 3S0 has been established by determination of the sulfur products and their oxidation states. Kinetic analysis leads to a four-step mechanism that accounts for the observed NO dependence. DFT calculations suggest the possibility that the nitrosylation product is a novel cluster [FeI4(NO)8(Cys)4]0 derived by dimerization of a pair of Roussin’s red ester (RRE) complexes.

AB - The reactivity of protein bound iron-sulfur clusters with nitric oxide (NO) is well documented, but little is known about the actual mechanism of cluster nitrosylation. Here, we report studies of members of the Wbl family of [4Fe-4S] containing proteins, which play key roles in regulating developmental processes in actinomycetes, including Streptomyces and Mycobacteria, and have been shown to be NO responsive. Streptomyces coelicolor WhiD and Mycobacterium tuberculosis WhiB1 react extremely rapidly with NO in a multiphasic reaction involving, remarkably, 8 NO molecules per [4Fe-4S] cluster. The reaction is 104-fold faster than that observed with O2 and is by far the most rapid iron-sulfur cluster nitrosylation reaction reported to date. An overall stoichiometry of [Fe4S4(Cys)4]2- + 8NO ? 2[FeI2(NO)4(Cys)2]0 + S2- + 3S0 has been established by determination of the sulfur products and their oxidation states. Kinetic analysis leads to a four-step mechanism that accounts for the observed NO dependence. DFT calculations suggest the possibility that the nitrosylation product is a novel cluster [FeI4(NO)8(Cys)4]0 derived by dimerization of a pair of Roussin’s red ester (RRE) complexes.

U2 - 10.1021/ja109581t

DO - 10.1021/ja109581t

M3 - Article

VL - 133

SP - 1112

EP - 1121

JO - Journal of the American Chemical Society

JF - Journal of the American Chemical Society

SN - 0002-7863

IS - 4

ER -