Buried S-nitrosocysteine revealed in crystal structures of human thioredoxin

Andrzej Weichsel, Jacqueline L. Brailey, William "Bill" Montfort

Research output: Contribution to journalArticle

54 Citations (Scopus)

Abstract

We have determined the 1.65 Å crystal structure of human thioredoxin-1 after treatment with S-nitrosoglutathione, providing a high-resolution view of this important protein modification and mechanistic insight into protein transnitrosation. Thioredoxin-1 appears to play an intermediary role in cellular S-nitrosylation and is important in numerous biological and pathobiological activities. S-Nitroso modifications of cysteines 62 and 69 are clearly visible in the structure and display planar cis geometries, whereas cysteines 32, 35, and 73 form intra- and intermolecular disulfide bonds. Surprisingly, the Cys 62 nitroso group is completely buried and pointing to the protein interior yet is the most readily formed at neutral pH. The Cys 69 nitroso group is also protected but requires a higher pH for stable formation. The helix intervening between residues 62 and 69 shifts by ∼0.5 Å to accommodate the SNO groups. The crystallographic asymmetric unit contains three independent molecules of thioredoxin, providing three views of the nitrosated protein. The three molecules are in general agreement but display subtle differences, including both cis and trans conformers for Cys 69 SNO in molecule C, and greater disorder in the Cys 62-Cys 69 helix in molecule B. Possible mechanisms for protein transnitrosation with specific geometric requirements and charge stabilization of the nitroxyl disulfide reaction intermediate are discussed.

Original languageEnglish (US)
Pages (from-to)1219-1227
Number of pages9
JournalBiochemistry
Volume46
Issue number5
DOIs
StatePublished - Feb 6 2007

Fingerprint

Thioredoxins
Crystal structure
Molecules
Proteins
Disulfides
Cysteine
S-Nitrosoglutathione
Reaction intermediates
Stabilization
S-nitrosocysteine
Geometry

ASJC Scopus subject areas

  • Biochemistry

Cite this

Buried S-nitrosocysteine revealed in crystal structures of human thioredoxin. / Weichsel, Andrzej; Brailey, Jacqueline L.; Montfort, William "Bill".

In: Biochemistry, Vol. 46, No. 5, 06.02.2007, p. 1219-1227.

Research output: Contribution to journalArticle

Weichsel, Andrzej ; Brailey, Jacqueline L. ; Montfort, William "Bill". / Buried S-nitrosocysteine revealed in crystal structures of human thioredoxin. In: Biochemistry. 2007 ; Vol. 46, No. 5. pp. 1219-1227.
@article{615c97bd1a4f4625842d55af69527705,
title = "Buried S-nitrosocysteine revealed in crystal structures of human thioredoxin",
abstract = "We have determined the 1.65 {\AA} crystal structure of human thioredoxin-1 after treatment with S-nitrosoglutathione, providing a high-resolution view of this important protein modification and mechanistic insight into protein transnitrosation. Thioredoxin-1 appears to play an intermediary role in cellular S-nitrosylation and is important in numerous biological and pathobiological activities. S-Nitroso modifications of cysteines 62 and 69 are clearly visible in the structure and display planar cis geometries, whereas cysteines 32, 35, and 73 form intra- and intermolecular disulfide bonds. Surprisingly, the Cys 62 nitroso group is completely buried and pointing to the protein interior yet is the most readily formed at neutral pH. The Cys 69 nitroso group is also protected but requires a higher pH for stable formation. The helix intervening between residues 62 and 69 shifts by ∼0.5 {\AA} to accommodate the SNO groups. The crystallographic asymmetric unit contains three independent molecules of thioredoxin, providing three views of the nitrosated protein. The three molecules are in general agreement but display subtle differences, including both cis and trans conformers for Cys 69 SNO in molecule C, and greater disorder in the Cys 62-Cys 69 helix in molecule B. Possible mechanisms for protein transnitrosation with specific geometric requirements and charge stabilization of the nitroxyl disulfide reaction intermediate are discussed.",
author = "Andrzej Weichsel and Brailey, {Jacqueline L.} and Montfort, {William {"}Bill{"}}",
year = "2007",
month = "2",
day = "6",
doi = "10.1021/bi061878r",
language = "English (US)",
volume = "46",
pages = "1219--1227",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "5",

}

TY - JOUR

T1 - Buried S-nitrosocysteine revealed in crystal structures of human thioredoxin

AU - Weichsel, Andrzej

AU - Brailey, Jacqueline L.

AU - Montfort, William "Bill"

PY - 2007/2/6

Y1 - 2007/2/6

N2 - We have determined the 1.65 Å crystal structure of human thioredoxin-1 after treatment with S-nitrosoglutathione, providing a high-resolution view of this important protein modification and mechanistic insight into protein transnitrosation. Thioredoxin-1 appears to play an intermediary role in cellular S-nitrosylation and is important in numerous biological and pathobiological activities. S-Nitroso modifications of cysteines 62 and 69 are clearly visible in the structure and display planar cis geometries, whereas cysteines 32, 35, and 73 form intra- and intermolecular disulfide bonds. Surprisingly, the Cys 62 nitroso group is completely buried and pointing to the protein interior yet is the most readily formed at neutral pH. The Cys 69 nitroso group is also protected but requires a higher pH for stable formation. The helix intervening between residues 62 and 69 shifts by ∼0.5 Å to accommodate the SNO groups. The crystallographic asymmetric unit contains three independent molecules of thioredoxin, providing three views of the nitrosated protein. The three molecules are in general agreement but display subtle differences, including both cis and trans conformers for Cys 69 SNO in molecule C, and greater disorder in the Cys 62-Cys 69 helix in molecule B. Possible mechanisms for protein transnitrosation with specific geometric requirements and charge stabilization of the nitroxyl disulfide reaction intermediate are discussed.

AB - We have determined the 1.65 Å crystal structure of human thioredoxin-1 after treatment with S-nitrosoglutathione, providing a high-resolution view of this important protein modification and mechanistic insight into protein transnitrosation. Thioredoxin-1 appears to play an intermediary role in cellular S-nitrosylation and is important in numerous biological and pathobiological activities. S-Nitroso modifications of cysteines 62 and 69 are clearly visible in the structure and display planar cis geometries, whereas cysteines 32, 35, and 73 form intra- and intermolecular disulfide bonds. Surprisingly, the Cys 62 nitroso group is completely buried and pointing to the protein interior yet is the most readily formed at neutral pH. The Cys 69 nitroso group is also protected but requires a higher pH for stable formation. The helix intervening between residues 62 and 69 shifts by ∼0.5 Å to accommodate the SNO groups. The crystallographic asymmetric unit contains three independent molecules of thioredoxin, providing three views of the nitrosated protein. The three molecules are in general agreement but display subtle differences, including both cis and trans conformers for Cys 69 SNO in molecule C, and greater disorder in the Cys 62-Cys 69 helix in molecule B. Possible mechanisms for protein transnitrosation with specific geometric requirements and charge stabilization of the nitroxyl disulfide reaction intermediate are discussed.

UR - http://www.scopus.com/inward/record.url?scp=33846783114&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33846783114&partnerID=8YFLogxK

U2 - 10.1021/bi061878r

DO - 10.1021/bi061878r

M3 - Article

C2 - 17260951

AN - SCOPUS:33846783114

VL - 46

SP - 1219

EP - 1227

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 5

ER -