In Solution Identification of the Lysine-Cysteine Redox Switch with a NOS Bridge in Transaldolase by Sulfur K-Edge X-ray Absorption Spectroscopy

J Phys Chem Lett. 2024 Apr 25;15(16):4263-4267. doi: 10.1021/acs.jpclett.4c00484. Epub 2024 Apr 12.

Abstract

A novel covalent post-translational modification (lysine-NOS-cysteine) was discovered in proteins, initially in the enzyme transaldolase of Neisseria gonorrhoeae (NgTAL) [Nature 2021, 593, 460-464], acting as a redox switch. The identification of this novel linkage in solution was unprecedented until now. We present detection of the NOS redox switch in solution using sulfur K-edge X-ray absorption spectroscopy (XAS). The oxidized NgTAL spectrum shows a distinct shoulder on the low-energy side of the rising edge, corresponding to a dipole-allowed transition from the sulfur 1s core to the unoccupied σ* orbital of the S-O group in the NOS bridge. This feature is absent in the XAS spectrum of reduced NgTAL, where Lys-NOS-Cys is absent. Our experimental and calculated XAS data support the presence of a NOS bridge in solution, thus potentially facilitating future studies on enzyme activity regulation mediated by the NOS redox switches, drug discovery, biocatalytic applications, and protein design.

MeSH terms

  • Cysteine / chemistry
  • Cysteine / metabolism
  • Lysine / chemistry
  • Lysine / metabolism
  • Neisseria gonorrhoeae / chemistry
  • Neisseria gonorrhoeae / enzymology
  • Oxidation-Reduction*
  • Protein Processing, Post-Translational
  • Solutions
  • Sulfur / chemistry
  • Sulfur / metabolism
  • Transaldolase* / chemistry
  • Transaldolase* / metabolism
  • X-Ray Absorption Spectroscopy*

Substances

  • Cysteine
  • Lysine
  • Solutions
  • Sulfur
  • Transaldolase