Professional Documents
Culture Documents
2010 Test1key PDF
2010 Test1key PDF
2 points
Chem 465
Biochemistry II
Multiple choice (4 points apiece):
4. Gluconeogenesis must use "bypass reactions" to circumvent three reactions in the glycolytic
pathway that are highly exergonic and essentially irreversible. Reactions carried out by which
three of the enzymes listed must be bypassed in the gluconeogenic pathway?
1) Hexokinase
2) Phosphoglycerate kinase
3) Phosphofructokinase-1
4) Pyruvate kinase
5) Triosephosphate isomerase
A) 1, 2, 3
B) 1, 2, 4
C) 1, 4, 5
D) 1, 3, 4
E) 2, 3, 4
5. Cellular isozymes of pyruvate kinase are allosterically inhibited by:
A) high concentrations of AMP.
B) high concentrations of ATP.
C) high concentrations of citrate.
D) low concentrations of acetyl-CoA.
E) low concentrations of ATP.
6. Which of the below is not required for the oxidative decarboxylation of pyruvate to form
acetyl-CoA?
A) ATP
B) CoA-SH
C) FAD
D) Lipoic acid
E) NAD+
7. Which one of the following enzymatic activities would be decreased by thiamine deficiency?
A) Fumarase
B) Isocitrate dehydrogenase
C) Malate dehydrogenase
D) Succinate dehydrogenase
E) á-Ketoglutarate dehydrogenase complex
(Page left blank so you can sketch out the glycolytic pathway to get your answers)
10. I am going to add glutamic acid labeled in the ä position with 13C (See diagram below) to a
cell that is using the TCA cycle to generate energy. Name, and draw the structure of the first
intermediate in the TCA cycle in which this label will appear. Indicate which C in this
intermediate will carry the 13C label. Initially glutamate will enter the TCA as á-ketoglutarate as labeled below:
Now draw the structures of these other TCA intermediates, and indicate where this label will
appear in these compounds
Succinyl-CoA Fumarate
Oxaloacetate Citrate
Note: due to symmetry in Succinate and Fumarate a single molecule of oxaloacetate will only
have a label at one COO- or the other, but not both. Similarly a single molecule of citrate will
have label at only one of the two positions indicated.
Isocitrate á-Ketoglutarate
Again only one of the labeled positions are labeled in any single isocitrate molecule.
Only ½ of the á-ketoglutarate molecules will be carrying a label after one turn of the TCA cycle.
(Page left blank so you can sketch out the TCA pathway to get your answers)
11. What is biotin? What is it used for biochemically? What enzymes have we studied that use
biotin as an integral part of their mechanism.
Biotin is a coenzyme or cofactor that carries activated CO2 in the active site of an enzyme. The
structure of biotin is shown below, but you were not required to know this structure.
12. Compare and contrast the structure, mechanism, cofactors used, and regulation of pyruvate
dehydrogenase complex and á-ketoglutarate dehydrogenase complex.
Pyruvate dehydrogenase and á-ketoglutarate dehydrogenase are very similar in structure and
mechanism. Both are large enzyme complexes that contain multiple copies of three major
proteins usually called simply E1, E2, and E3. Both enzyme complexes also utilize the cofactors
thiamine pyrophosphate, lippoate and FAD, ad well as the substrate CoenzymeA.
In pyruvate dehydrogenase the E1 protein is actually the pyruvate dehydrogenase part of the
complex. It uses TPP to cleave the terminal CO2 from pyruvate and transfers the remaining part
of the pyruvate to E2 (Dihydrolipoyl transacetylase) where this acetate group is temporarily
attached to lippoate. The acetate is then transferred to free CoASH to make acetyl CoA which
diffuses away from the enzyme complex. When this transfer occurs the lipoate is left in a fully
reduced form that must be regenerated before the enzyme can work again. This oxidation is
performed by the E3 enzyme, dihydrolipoyl dehydrogenase, which transfers the protons from the
reduced lipoate to NAD+ to make NADH.
The E3 enzyme is virtually identical in both complexes because it does exactly the same
biochemical reaction. The E1 enzyme is just a bit different in the two complexes because it must
bind different substrates (pyruvate or á-ketoglutarate). The E2 complexes are close in structure
because they also do the same chemistry, but in they vary slightly because different one creates
acetyl CoA and the other makes sussinyl-CoA.
The allosteric control of pyruvate dehydrogenase is a bit more complicated because it is the first
step in the TCA pathway. Pyruvate dehydrogenase is stimulated AMP, CoA, NAD+, and Ca2+
and inhibited by ATP, acetyl-CoA, NADH. The á-ketoglutarate dehydrogenase control is
simpler because it is deeper inside the TCA cycle. It is only stimulated by Ca2+ and inhibited by
succinyl-CoA and NADH.
1: C
2: E
3: A
4: D
5: B
6: A
7: E