Reversible S-nitrosylation in an engineered azurin

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

doi:10.1038/nchem.2489

Reversible S-nitrosylation in an engineered azurin

Journal Article · Mon Apr 25 00:00:00 EDT 2016 · Nature Chemistry

DOI:https://doi.org/10.1038/nchem.2489· OSTI ID:1340343

Tian, Shiliang ^[1]; Liu, Jing ^[1]; Cowley, Ryan E. ^[2]; Hosseinzadeh, Parisa ^[1]; Marshall, Nicholas M. ^[1]; ^[1]; Robinson, Howard ^[3]; Nilges, Mark J. ^[1]; Blackburn, Ninian J. ^[4]; Solomon, Edward I. ^[2]; Lu, Yi ^[1]

Univ. of Illinois at Urbana-Champaign, Urbana, IL (United States)
Stanford Univ., Stanford, CA (United States)
Brookhaven National Lab. (BNL), Upton, NY (United States)
Oregon Health & Sciences Univ., Portland, OR (United States)

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.

View Accepted Manuscript (DOE)

Research Organization:: Brookhaven National Laboratory (BNL), Upton, NY (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)

OSTI ID:: 1340343

Alternate ID(s):: OSTI ID: 1351345

Report Number(s):: BNL--112002-2016-JA

Journal Information:: Nature Chemistry, Journal Name: Nature Chemistry Journal Issue: 7 Vol. 8; ISSN 1755-4330

Publisher:: Nature Publishing GroupCopyright Statement

Country of Publication:: United States

Language:: English

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