Determining the quantitative relationship between glycolysis and GAPDH in cancer cells exhibiting the Warburg effect

Autor: Chengmeng Jin, Qiangrong Pan, Xun Hu, Xiaobing Zhu
Rok vydání: 2021
Předmět:
0301 basic medicine
F6P
fructose 6-phosphate
DHAP
dihydroxyacetone phosphate
Biochemistry
NC
negative control
chemistry.chemical_compound
cell metabolism
Neoplasms
Warburg Effect
Oncologic
RNA
Small Interfering
Phosphoglycerate kinase 1
Glyceraldehyde 3-phosphate dehydrogenase
PGAM
phosphoglycerate mutase
education.field_of_study
biology
GAPDH-SA
GAPDH specific activity
LDH
lactate dehydrogenase
GAPDH
Glyceraldehyde-3-Phosphate Dehydrogenases
glycolysis
Glc
glucose
Warburg effect
TPI
triose phosphate isomerase
GA3P
glyceraldehyde 3-phosphate
flux control
Gibbs free energy
Oxidation-Reduction
Research Article
FBP
fructose 1
6-bisphosphate
G6P
glucose 6-phosphate
03 medical and health sciences
stomatognathic system
FBS
fetal bovine serum
G6PDH
glucose-6-phosphate dehydrogenase
Cell Line
Tumor
PGI
phosphohexose isomerase
PGK
phosphoglycerate kinase
Humans
IA
iodoacetate
education
Molecular Biology
GAPDH-AA
GAPDH actual activity
Dihydroxyacetone phosphate
UPLC
ultra performance liquid chromatography
Hexokinase
030102 biochemistry & molecular biology
PEP
phosphoenolpyruvate
Cell Biology
HK
hexokinase
Pyr
pyruvate
030104 developmental biology
Glucose
chemistry
Glucose 6-phosphate
Anaerobic glycolysis
PK
pyruvate kinase
biology.protein
cancer cells
WE
Warburg effect
Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating)
Pyruvate kinase
Zdroj: The Journal of Biological Chemistry
ISSN: 1083-351X
Popis: Previous studies have identified GAPDH as a promising target for treating cancer and modulating immunity because its inhibition reduces glycolysis in cells (cancer cells and immune cells) with the Warburg effect, a modified form of cellular metabolism found in cancer cells. However, the quantitative relationship between GAPDH and the aerobic glycolysis remains unknown. Here, using siRNA-mediated knockdown of GAPDH expression and iodoacetate-dependent inhibition of enzyme activity, we examined the quantitative relationship between GAPDH activity and glycolysis rate. We found that glycolytic rates were unaffected by the reduction of GAPDH activity down to 19% ± 4.8% relative to untreated controls. However, further reduction of GAPDH activity below this level caused proportional reductions in the glycolysis rate. GAPDH knockdown or inhibition also simultaneously increased the concentration of glyceraldehyde 3-phosphate (GA3P, the substrate of GAPDH). This increased GA3P concentration countered the effect of GAPDH knockdown or inhibition and stabilized the glycolysis rate by promoting GAPDH activity. Mechanistically, the intracellular GA3P concentration is controlled by the Gibbs free energy of the reactions upstream of GAPDH. The thermodynamic state of the reactions along the glycolysis pathway was only affected when GAPDH activity was reduced below 19% ± 4.8%. Doing so moved the reactions catalyzed by GAPDH + PGK1 (phosphoglycerate kinase 1, the enzyme immediate downstream of GAPDH) away from the near-equilibrium state, revealing an important biochemical basis to interpret the rate control of glycolysis by GAPDH. Collectively, we resolved the numerical relationship between GAPDH and glycolysis in cancer cells with the Warburg effect and interpreted the underlying mechanism.
Databáze: OpenAIRE
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