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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