Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2025 Nov:706:115944.
doi: 10.1016/j.ab.2025.115944. Epub 2025 Jul 11.

Spectroscopic method for measuring activity of cis-aconitate decarboxylase, an important metabolic regulator of immune responses

Kevin Knowlan  1 , Cody L Hoop  2 , Nadya I Tarasova  3
Affiliations

Spectroscopic method for measuring activity of cis-aconitate decarboxylase, an important metabolic regulator of immune responses

Kevin Knowlan et al. Anal Biochem. 2025 Nov.

Abstract

Cis-aconitate decarboxylase (ACOD1) is a key enzyme converting cis-aconitate to itaconate, which has therapeutic potential for inflammatory diseases. Existing methods to measure ACOD1 activity and itaconate are often expensive and complex. We developed a novel, high-throughput spectrophotometric assay using the Fürth-Herrmann reaction. Our method quantifies ACOD1-catalyzed itaconate production by leveraging distinct absorbance ratios of cis-aconitate and itaconate at 386 nm and 440 nm. We optimized parameters, characterized human ACOD1 kinetics, and determined an IC50 for citraconate consistent with previous reports. This simple, fast, and reliable assay, requiring only a UV-Vis spectrophotometer, will accelerate screening for ACOD1 modulators, speeding up therapeutic development.

Keywords: Immune-Responsive Gene 1 (IRG1); Itaconate; Spectrophotometry; cis-aconitate decarboxylase (ACOD1).

PubMed Disclaimer

Conflict of interest statement

Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

References

    1. Qian C, Wang Y, Yuan Q, Guo Y, Wang Y, Insights into the itaconate family: Immunomodulatory mechanisms and therapeutic potentials, Eur J Pharmacol, 997 (2025) 177542. - PubMed
    1. Williams NC, O’Neill LAJ, A Role for the Krebs Cycle Intermediate Citrate in Metabolic Reprogramming in Innate Immunity and Inflammation, Front Immunol, 9 (2018). - PMC - PubMed
    1. Wu R, Kang R, Tang D, Mitochondrial ACOD1/IRG1 in infection and sterile inflammation, J Intensive Med, 2 (2022) 78–88. - PMC - PubMed
    1. Lampropoulou V, Sergushichev A, Bambouskova M, Nair S, Vincent EE, Loginicheva E, Cervantes-Barragan L, Ma X, Huang SC, Griss T, Weinheimer CJ, Khader S, Randolph GJ, Pearce EJ, Jones RG, Diwan A, Diamond MS, Artyomov MN, Itaconate Links Inhibition of Succinate Dehydrogenase with Macrophage Metabolic Remodeling and Regulation of Inflammation, Cell Metab, 24 (2016) 158–166. - PMC - PubMed
    1. Sakai A, Kusumoto A, Kiso Y, Furuya E, Itaconate reduces visceral fat by inhibiting fructose 2,6-bisphosphate synthesis in rat liver, Nutrition, 20 (2004) 997–1002. - PubMed

LinkOut - more resources