Crystal structures of reduced, oxidized, and mutated human thioredoxins

Evidence for a regulatory homodimer

Andrzej Weichsel, John R. Gasdaska, Garth Powis, William "Bill" Montfort

Research output: Contribution to journalArticle

286 Citations (Scopus)

Abstract

Background: Human thioredoxin reduces the disulfide bonds of numerous proteins in vitro, and can activate transcription factors such as NFκB in vivo. Thioredoxin can also act as a growth factor, and is overexpressed and secreted in certain tumor cells. Results: Crystal structures were determined for reduced and oxidized wild type human thioredoxm (at 1.7 and 2.1 Å nominal resolution, respectively), and for reduced mutant proteins Cys73→Ser and Cys32→Ser/Cys35→Ser (at 1.65 and 1.8 Å, respectively), Surprisingly, thioedoxin is dimeric in all four structures; the dimer is linked through a disulfide bond between Cys73 of each monomer, except in Cys73→Ser where a hydrogen bond occurs. The thioredoxin active site is blocked by dimer formation. Conformational changes in the active site and dimer interface accompany oxidation of the active-site cysteines, Cys32 and Cys35.ts Conclusions: It has been suggested that a reduced Ka in the first cysteine (Cys32 in human thioredoxin) of the active-site sequence is important for modulation of the redox potential in thioredoxin. A hydrogen bond between the sulfhydryls of Cys32 and Cys35 may reduce the pKa of Cys32 and this pKa depression probably results in increased nucleophilicity of the Cys32 thiolate group. This nucleophilicity, in turn, is thought to be necessary for the role of thioredoxin in disufide-bond reduction. The physiological role, if any of thioredoxin dimer formation remains unknown. It is possible that dimerization may provide a mechanism for regulation of the protein, or a means of sensing oxidative stress.

Original languageEnglish (US)
Pages (from-to)735-751
Number of pages17
JournalStructure
Volume4
Issue number6
StatePublished - 1996

Fingerprint

Thioredoxins
Catalytic Domain
Disulfides
Cysteine
Hydrogen
Dimerization
Mutant Proteins
Oxidation-Reduction
Intercellular Signaling Peptides and Proteins
Proteins
Oxidative Stress
Transcription Factors

Keywords

  • Dimer
  • Mutant
  • Oxidation
  • Thioredoxin
  • X-ray crystallography

ASJC Scopus subject areas

  • Molecular Biology
  • Structural Biology

Cite this

Crystal structures of reduced, oxidized, and mutated human thioredoxins : Evidence for a regulatory homodimer. / Weichsel, Andrzej; Gasdaska, John R.; Powis, Garth; Montfort, William "Bill".

In: Structure, Vol. 4, No. 6, 1996, p. 735-751.

Research output: Contribution to journalArticle

Weichsel, Andrzej ; Gasdaska, John R. ; Powis, Garth ; Montfort, William "Bill". / Crystal structures of reduced, oxidized, and mutated human thioredoxins : Evidence for a regulatory homodimer. In: Structure. 1996 ; Vol. 4, No. 6. pp. 735-751.
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