Analysis of GAPDH and Protein Interaction

Ganapathy-Kanniappan, Shanmugasundaram

doi:10.1007/978-981-10-7342-7_5

Cited by 2 publications

(3 citation statements)

References 17 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…NADH channeling between byGAPDH and mammalian LDH isozymes can be attributed to highly conserved structures in GAPDH family of enzymes 6 . The conserved structure could be a result of evolutionary pressure to conserve the related interactome 6,11,27 .…”

Section: Discussionmentioning

confidence: 99%

“…The changes in NAD+/NADH concentrations have a peculiarly strong effect on GAPDH structure 1,2,6,23-26 . A decrease in NADH concentration leads to partial dissociation of the GAPDH tetramers 2,6,27 . NAD(H) binding to different subunits in GAPDH tetramer results in allosteric regulation of GAPDH activity and strong negative cooperativity that can change the NAD(H) binding affinity by thousand-fold 24-26 .…”

Section: Introductionmentioning

confidence: 99%

“…NAD(H) binding to different subunits in GAPDH tetramer results in allosteric regulation of GAPDH activity and strong negative cooperativity that can change the NAD(H) binding affinity by thousand-fold 24-26 . These effects are species specific and known for decades, but their physiological significance is still unknown 2,6,27 .…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Substrate Channelingviaa Transient Protein-Protein Complex: The case of D-Glyceraldehyde-3-Phosphate Dehydrogenase and L-Lactate Dehydrogenase

Svedružić

Odorčić

Chang

et al. 2020

Preprint

View full text Add to dashboard Cite

Background: D-Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and L-lactate dehydrogenase (LDH) can form a complex that can regulate the major metabolic pathways, however, the exact mechanism remains unknown. We analyzed a possibility of NADHchanneling from GAPDH-NADH complex to LDH isozymes using enzymes from different cells. Results: Enzyme-kinetics and NADH-binding studies showed that LDH can use GAPDH-NADH complex as a substrate. LDH activity with GAPDH-NADH complex was challenged with anti-LDH antibodies to show that the channeled and the diffusive reactions always take place in parallel. The channeling path is dominant only in assays with limiting free-NADH concertation that mimic cytosolic conditions. Analytical ultracentrifugation showed that the channeling does not require a high affinity complex. Molecular dynamics calculations and protein-protein interaction studies showed that LDH and GAPDH can form a leaky channeling complex only at subsaturating NADH concentrations. The interaction sites are conserved between LDH isozymes from heart and muscle, and between GAPDH molecules from rabbit and yeast cells. Positive electric fields between the NAD(H) binding sites on LDH and GAPDH tetramers, showed that NAD(H)-channeling within the LDH-GAPDH complex, can be an extension of NAD(H)channeling between the adjacent subunits in each tetramer. Conclusions: In the case of a transient (GAPDH-NADH)-LDH complex, the relative contribution from the channeled and the diffusive paths depends on the overlap between off-rates for the transient (GAPDH-NADH)-LDH complex and off-rates for the GAPDH-NADH complex. Molecular evolution or metabolic engineering protocols can exploit substrate channeling for metabolic flux control by fine-tuning substrate-binding affinity for the key enzymes in the competing reaction paths. Highlights-Substrate channeling molecular mechanism can regulate energy production and aerobic and anaerobic metabolism in cells -LDH and GAPDH can form a channeling complex only at sub-saturating NADH concentration -Channeled and diffusive paths always compete and take place in parallel -NADH channeling does not require a high-affinity complex -NADH channeling within GAPDH-LDH complex is an extension of NAD(H) channeling within each tetramer -Allosteric modulation of NADH binding affinity in GAPDH tetramer can regulate NAD(H) channeling Introduction D-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and L-lactate dehydrogenases (LDH) are two NAD(H) dependent dehydrogenases that participate in glycolytic and gluconeogenic pathways 1,2 . Glycolysis and gluconeogenesis are the two major metabolic pathways that provide cells with metabolic precursors and a rapid source of energy. Parallel to glycolysis GAPDH participates in microtubule bundling, DNA replication and repair, apoptosis, the export of nuclear RNA, membrane fusion, and phosphotransferase activity 1,2 . GAPDH is implicated in Huntington's disease 3 , prostate cancer, and viral pathogenesis 1,2 . GAPDH could be a target of nitric oxide 4 and a target of ...

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Substrate Channelingviaa Transient Protein-Protein Complex: The case of D-Glyceraldehyde-3-Phosphate Dehydrogenase and L-Lactate Dehydrogenase

Svedružić

Odorčić

Chang

et al. 2020

Preprint

View full text Add to dashboard Cite

show abstract

Substrate Channeling via a Transient Protein-Protein Complex: The case of D-Glyceraldehyde-3-Phosphate Dehydrogenase and L-Lactate Dehydrogenase

Svedružić

Odorčić

Chang

et al. 2020

Sci Rep

View full text Add to dashboard Cite

Substrate channeling studies have frequently failed to provide conclusive results due to poor understanding of this subtle phenomenon. We analyzed the mechanism of NADH-channeling from D-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) to L-lactate Dehydrogenase (LDH) using enzymes from different cells. Enzyme kinetics studies showed that LDH activity with free NADH and GAPDH-NADH complex always take place in parallel. The channeling is observed only in assays that mimic cytosolic conditions where free NADH concentration is negligible and the GAPDH-NADH complex is dominant. Molecular dynamics and protein-protein interaction studies showed that LDH and GAPDH can form a leaky channeling complex only at the limiting NADH concentrations. Surface calculations showed that positive electric field between the NAD(H) binding sites on LDH and GAPDH tetramers can merge in the LDH-GAPDH complex. NAD(H)-channeling within the LDH-GAPDH complex can be an extension of NAD(H)-channeling within each tetramer. In the case of a transient LDH-(GAPDH-NADH) complex, the relative contribution from the channeled and the diffusive paths depends on the overlap between the off-rates for the LDH-(GAPDH-NADH) complex and the GAPDH-NADH complex. Molecular evolution or metabolic engineering protocols can exploit substrate channeling for metabolic flux control by fine-tuning substrate-binding affinity for the key enzymes in the competing reaction paths.

show abstract

Analysis of GAPDH and Protein Interaction

Cited by 2 publications

References 17 publications

Substrate Channelingviaa Transient Protein-Protein Complex: The case of D-Glyceraldehyde-3-Phosphate Dehydrogenase and L-Lactate Dehydrogenase

Substrate Channelingviaa Transient Protein-Protein Complex: The case of D-Glyceraldehyde-3-Phosphate Dehydrogenase and L-Lactate Dehydrogenase

Substrate Channeling via a Transient Protein-Protein Complex: The case of D-Glyceraldehyde-3-Phosphate Dehydrogenase and L-Lactate Dehydrogenase

Contact Info

Product

Resources

About