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Title: Reversible S-nitrosylation in an engineered azurin

Here, S-Nitrosothiols are known as reagents for NO storage and transportation and as regulators in many physiological processes. Although the S-nitrosylation catalysed by haem proteins is well known, no direct evidence of S-nitrosylation in copper proteins has been reported. Here, we report reversible insertion of NO into a copper–thiolate bond in an engineered copper centre in Pseudomonas aeruginosa azurin by rational design of the primary coordination sphere and tuning its reduction potential by deleting a hydrogen bond in the secondary coordination sphere. The results not only provide the first direct evidence of S-nitrosylation of Cu(II)-bound cysteine in metalloproteins, but also shed light on the reaction mechanism and structural features responsible for stabilizing the elusive Cu(I)–S(Cys)NO species. The fast, efficient and reversible S-nitrosylation reaction is used to demonstrate its ability to prevent NO inhibition of cytochrome bo 3 oxidase activity by competing for NO binding with the native enzyme under physiologically relevant conditions.
Authors:
 [1] ;  [1] ;  [2] ;  [1] ;  [1] ; ORCiD logo [1] ;  [3] ;  [1] ;  [4] ;  [2] ;  [1]
  1. Univ. of Illinois at Urbana-Champaign, Urbana, IL (United States)
  2. Stanford Univ., Stanford, CA (United States)
  3. Brookhaven National Lab. (BNL), Upton, NY (United States)
  4. Oregon Health & Sciences Univ., Portland, OR (United States)
Publication Date:
Report Number(s):
BNL-112002-2016-JA
Journal ID: ISSN 1755-4330
Grant/Contract Number:
SC00112704
Type:
Accepted Manuscript
Journal Name:
Nature Chemistry
Additional Journal Information:
Journal Volume: 8; Journal Issue: 7; Journal ID: ISSN 1755-4330
Publisher:
Nature Publishing Group
Research Org:
Brookhaven National Laboratory (BNL), Upton, NY (United States)
Sponsoring Org:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)
Country of Publication:
United States
Language:
English
Subject:
36 MATERIALS SCIENCE
OSTI Identifier:
1340343

Tian, Shiliang, Liu, Jing, Cowley, Ryan E., Hosseinzadeh, Parisa, Marshall, Nicholas M., Yu, Yang, Robinson, Howard, Nilges, Mark J., Blackburn, Ninian J., Solomon, Edward I., and Lu, Yi. Reversible S-nitrosylation in an engineered azurin. United States: N. p., Web. doi:10.1038/nchem.2489.
Tian, Shiliang, Liu, Jing, Cowley, Ryan E., Hosseinzadeh, Parisa, Marshall, Nicholas M., Yu, Yang, Robinson, Howard, Nilges, Mark J., Blackburn, Ninian J., Solomon, Edward I., & Lu, Yi. Reversible S-nitrosylation in an engineered azurin. United States. doi:10.1038/nchem.2489.
Tian, Shiliang, Liu, Jing, Cowley, Ryan E., Hosseinzadeh, Parisa, Marshall, Nicholas M., Yu, Yang, Robinson, Howard, Nilges, Mark J., Blackburn, Ninian J., Solomon, Edward I., and Lu, Yi. 2016. "Reversible S-nitrosylation in an engineered azurin". United States. doi:10.1038/nchem.2489. https://www.osti.gov/servlets/purl/1340343.
@article{osti_1340343,
title = {Reversible S-nitrosylation in an engineered azurin},
author = {Tian, Shiliang and Liu, Jing and Cowley, Ryan E. and Hosseinzadeh, Parisa and Marshall, Nicholas M. and Yu, Yang and Robinson, Howard and Nilges, Mark J. and Blackburn, Ninian J. and Solomon, Edward I. and Lu, Yi},
abstractNote = {Here, S-Nitrosothiols are known as reagents for NO storage and transportation and as regulators in many physiological processes. Although the S-nitrosylation catalysed by haem proteins is well known, no direct evidence of S-nitrosylation in copper proteins has been reported. Here, we report reversible insertion of NO into a copper–thiolate bond in an engineered copper centre in Pseudomonas aeruginosa azurin by rational design of the primary coordination sphere and tuning its reduction potential by deleting a hydrogen bond in the secondary coordination sphere. The results not only provide the first direct evidence of S-nitrosylation of Cu(II)-bound cysteine in metalloproteins, but also shed light on the reaction mechanism and structural features responsible for stabilizing the elusive Cu(I)–S(Cys)NO species. The fast, efficient and reversible S-nitrosylation reaction is used to demonstrate its ability to prevent NO inhibition of cytochrome bo3 oxidase activity by competing for NO binding with the native enzyme under physiologically relevant conditions.},
doi = {10.1038/nchem.2489},
journal = {Nature Chemistry},
number = 7,
volume = 8,
place = {United States},
year = {2016},
month = {4}
}