Monothiol and dithiol glutaredoxin-1 from Clostridium oremlandii: Identification of domain-swapped structures by NMR, X-ray crystallography and HDX mass spectrometry

Kitaik Lee, Kwon Joo Yeo, Sae Hae Choi, Eun Hye Lee, Bo Keun Kim, Sulhee Kim, Hae Kap Cheong, Won Kyu Lee, Hwa Young Kim, Eunha Hwang, Ju Rang Woo, Sung Joon Lee, Kwang Yeon Hwanga

Research output: Contribution to journalArticlepeer-review

Abstract

Protein dimerization or oligomerization resulting from swapping part of the protein between neighboring polypeptide chains is known to play a key role in the regulation of protein function and in the formation of protein aggregates. Glutaredoxin-1 from Clostridium oremlandii (cGrx1) was used as a model to explore the formation of multiple domain-swapped conformations, which were made possible by modulating several hinge-loop residues that can form a pivot for domain swapping. Specifically, two alternative domain-swapped structures were generated and analyzed using nuclear magnetic resonance (NMR), X-ray crystallography, circular-dichroism spectroscopy and hydrogen/deuterium-exchange (HDX) mass spectrometry. The first domain-swapped structure (β3-swap) was formed by the hexameric cGrx1-cMsrA complex. The second domain-swapped structure (β1-swap) was formed by monothiol cGrx1 (C16S) alone. In summary, the first domain-swapped structure of an oxidoreductase in a hetero-oligomeric complex is presented. In particular, a single point mutation of a key cysteine residue to serine led to the formation of an intramolecular disulfide bond, as opposed to an intermolecular disulfide bond, and resulted in modulation of the underlying free-energy landscape of protein oligomerization.

Original languageEnglish
Pages (from-to)1019-1027
Number of pages9
JournalIUCrJ
Volume7
DOIs
Publication statusPublished - 2020 Nov 1

Keywords

  • disulfide bonds
  • domain swapping
  • glutaredoxin
  • oxidoreductases

ASJC Scopus subject areas

  • Chemistry(all)
  • Biochemistry
  • Materials Science(all)
  • Condensed Matter Physics

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