Human hexokinase 2 is an essential regulator of glycolysis that couples metabolic and pro-
liferative activities in cancer cells. The binding of hexokinase 2 to the outer membrane of
mitochondria is critical for its oncogenic activity. However, the regulation of hexokinase 2
binding to mitochondria remains unclear. Here, we report that SUMOylation regulates the
binding of hexokinase 2 to mitochondria. We find that hexokinase 2 can be SUMOylated at
K315 and K492. SUMO-specific protease SENP1 mediates the de-SUMOylation of hexokinase
2. SUMO-defective hexokinase 2 preferably binds to mitochondria and enhances both glu-
cose consumption and lactate production and decreases mitochondrial respiration in parallel.
This metabolic reprogramming supports prostate cancer cell proliferation and protects cells
from chemotherapy-induced cell apoptosis. Moreover, we demonstrate an inverse relation-
ship between SENP1-hexokinase 2 axis and chemotherapy response in prostate cancer
samples. Our data provide evidence for a previously uncovered posttranslational modification
of hexokinase 2 in cancer cells, suggesting a potentially actionable strategy for preventing
chemotherapy resistance in prostate cancer.
