Research Paper 1 Flashcards
What is the goal of this study?
Identify a minimal target site that can be used for the development of a selective GAPDH inhibitor. They want to find a GAPDH mutation that will disrupt glycolysis.
Based on the introduction and the discussion, what was previously known about
the relationship between GAPDH and cancer?
GAPDH has increased activity in cancer cells, and blocking its activity with inhibitors or antagonists has been observed to block cancer progression.
What region of GAPDH do the authors decide to mutate? What region of the
protein do the authors specifically avoid mutating?
They want to mutate the NAD+ binding region and avoid mutating the substrate binding region where Glyceraldehyde 3 phosphate binds.
. In Figure 2, how does the mutant GAPDH impact the function of wild-type
(unmutated) GAPDH? (
The mutant GAPDH competitively inhibited wild-type GAPDH.
What is the effect of the GAPDH mutation on glucose uptake by cells as seen in
Figure 3A?
The GAPDH mutation results in decreased glucose uptake.
Mutation of GAPDH would be expected to disrupt glycolysis leading to cellular
stress.
What proteins were examined by immunoblotting in Figure 3B to assess if
there was an increase in cellular stress?
What was observed?
phosphorylated AKT and AMPK were the proteins examined to determine if cellular stress increased. The levels of these proteins increased, showing that cellular stress was increased.
How does the ectopic expression of GAPDH affect the % of metabolically active
cells, as shown in Figure 4?
Ectopic expression of GAPDH decreased the percent of metabolically active cells
Based on the results in Figure 5, what effect does the GAPDH mutation have on
cell growth and viability?
What protein is examined to observe the effects on cell proliferation and what is the effect of mutant GAPDH on cell proliferation?
What protein is observed to examine levels of protein synthesis and what do they
conclude about the effects of mutant GAPDH on the levels of protein synthesis?
GAPDH mutation causes decrease in cell viability and cell growth.
To look at cell proliferation the author examined the levels of PCNA by Western blotting and observed a decrease in PCNA in the presence of mutant GAPDH. T
To examine translation levels, authors observed levels of p-elf2a and observed an increase in levels, showing a decrease in translation.
What effect does mutant GAPDH have on tumor progression as shown in Figure
6?
Mutant GAPDH decreases tumor progression.
