Alternol inhibits GAPDH activity and disrupts glycolytic flux preferentially in cancer cells
Year: 2026
Type: article
Abstract: Introduction We and others demonstrated that the natural compound Alternol induces apoptosis preferentially in human cancer cells. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) participates in cellular glycolysis, important for energy homeostasis, especially in cancer cells. We recently discovered that Alternol interacts with GAPDH, along with 4 Krebs cycle enzymes. In this study, we characterized the mechanism for Alternol-GAPDH interaction and the functional significance. Methods Multiple human prostate cancer cell lines and a benign prostate epithelial cell line were utilized in the experiments. Enzyme activity assay in vitro with purified protein was used to examine Alternol inhibition of GAPDH activity. Computer-based docking assessment was performed to analyze Alternol interaction with GAPDH protein. Seahorse instrument was used to conduct glycolytic testing. Results Our data revealed that Alternol interacts with GAPDH protein on two sites, one of which is the NAD + binding site on the active domain of the enzyme, postulating an inhibitory effect. As expected, Alternol directly inhibited GAPDH dehydrogenase activity in an in vitro assay with purified enzyme with nanomole IC50 value at 5.794 nM. Consistently, Alternol significantly suppressed its enzymatic activity in cultured cancer cells but not in benign cells. These inhibitory effects were associated with reduced glycolytic capacity in cancer cells as assessed by extracellular acidification rate (ECAR) and metabolomic analysis. Conclusion These results suggest that Alternol potently inhibits GAPDH activity and specifically disrupts glycolytic flux in cancer cells.
Source: Frontiers in Pharmacology
Authors: Yuelong Chai, Wang Liu, Moben Mirza, Hua Chen, Benyi Li
Language: English
Subfield: Cancer Research
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $3,801
OpenAlex ID: W7165787002