Medicinal Chemistry & Chemical Biology, Short talk
MC-016

Toward a Protein-Based Sensor for S-Nitrosylated Coenzyme A

H. Lämmermann1, P. Rivera Fuentes1*
1Department of Chemistry, University of Zurich, Winterthurerstrasse 190, 8057 Zurich, Switzerland

The S-nitrosylation of protein cysteine residues, resulting in the formation of S-nitrosothiols (SNOs), has emerged as an important signaling pathway of nitric oxide (NO).[1] This well-regulated post-translational modification plays a role in protein activation or inhibition, as well as the modulation of protein-protein interactions and protein localization.[2] Recently, S-nitrosylated coenzyme A (SNO-CoA) was identified as a key mediator of NO transfer.[3,4]

To investigate SNO-CoA signaling, we aim to develop a genetically encoded SNO-CoA sensor based on the fusion of redox-sensitive green fluorescent protein (roGFP)[5] with a CoA-binding protein (Figure 1). In this design, NO is transferred from the CoA-binding domain to the cysteine residues of roGFP, producing a ratiometric fluorescence change. To achieve efficient and selective NO transfer, we employ de novo protein design to engineer a structured linker that enforces the precise spatial orientation between the CoA-binding domain and roGFP.

Figure 1. Fusion Protein Design. In green roGFP with cysteines represented as sticks, in grey CoA-binding protein with SNO-CoA bound.

This approach will enable measurements of SNO-CoA in live cells, providing a direct readout of the S-nitrosylation state of CoA and offering new insights into this emerging signaling pathway.

[1] Veani Fernando, Xunzhen Zheng, Yashna Walia, Vandana Sharma, Joshua Letson, Saori Furuta, Antioxidants 2019, 8, 404.
[2] Douglas T. Hess, Akio Matsumoto, Sung-Oog Kim, Harvey E. Marshall, Jonathan S. Stamler, Nat. Rev. Mol. Cell Biol. 2005, 6, 150–166.
[3] Puneet Anand, Alfred Hausladen, Ya-Juan Wang, Guo-Fang Zhang, Colin Stomberski, Henri Brunengraber, Douglas T. Hess, Jonathan S. Stamler, Proc. Natl. Acad. Sci. U.S.A. 2014, 111, 18572–18577.
[4] Hua-Lin Zhou, Zachary W. Grimmett, Nicholas M. Venetos, Colin T. Stomberski, Zhaoxia Qian, Precious J. McLaughlin, Puneet K. Bansal, Rongli Zhang, James D. Reynolds, Richard T. Premont, Jonathan S. Stamler, Cell 2023, 186, 5812.
[5] George T. Hanson, Robert Aggeler, Devin Oglesbee, Mark Cannon, Roderick A. Capaldi, Roger Y. Tsien, S. James Remington, J. Biol. Chem.2004, 279, 13044.