American Association for Cancer Research
00085472can151023-sup-148116_1_supp_3084931_nshg19.doc (971 kB)

Supplementary information from JX06 Selectively Inhibits Pyruvate Dehydrogenase Kinase PDK1 by a Covalent Cysteine Modification

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journal contribution
posted on 2023-03-31, 00:00 authored by Wenyi Sun, Zuoquan Xie, Yifu Liu, Dan Zhao, Zhixiang Wu, Dadong Zhang, Hao Lv, Shuai Tang, Nan Jin, Hualiang Jiang, Minjia Tan, Jian Ding, Cheng Luo, Jian Li, Min Huang, Meiyu Geng

Supplementary experimental procedures and figure legend



Pyruvate dehydrogenase kinase PDK1 is a metabolic enzyme responsible for switching glucose metabolism from mitochondrial oxidation to aerobic glycolysis in cancer cells, a general hallmark of malignancy termed the Warburg effect. Herein we report the identification of JX06 as a selective covalent inhibitor of PDK1 in cells. JX06 forms a disulfide bond with the thiol group of a conserved cysteine residue (C240) based on recognition of a hydrophobic pocket adjacent to the ATP pocket of the PDK1 enzyme. Our investigations of JX06 mechanism suggested that covalent modification at C240 induced conformational changes at Arginine 286 through Van der Waals forces, thereby hindering access of ATP to its binding pocket and in turn impairing PDK1 enzymatic activity. Notably, cells with a higher dependency on glycolysis were more sensitive to PDK1 inhibition, reflecting a metabolic shift that promoted cellular oxidative stress and apoptosis. Our findings offer new mechanistic insights including how to therapeutically target PDK1 by covalently modifying the C240 residue. Cancer Res; 75(22); 4923–36. ©2015 AACR.