CHEM 464 Fall 2011

Final Exam 12/15/11

Student Name__________________________

Instructions:
1. You must use Scantron Form 815-E to answer Question 1 (Multiple Choice).
Please be sure to return the Scantron form separately from the exam.
2. For all other questions, write answers in the space provided; use reverse side
if necessary.
3. Use of a non-programmable calculator is permitted.
4. pKa of amino acid functional groups, and Standard Reduction Potentials of
electron carriers are provided on the last page. You may separate this page
from the exam; you dont need to return this page with your exam.
5. Please show calculations used to solve problems. This will enable you to earn
partial credit even if your final answer is incorrect.

2. Provide short answers to each of the following questions (a through h):

a) In what order would dipeptides Arg-Lys, His-Lys, and Leu-tyr be eluted from a carboxymethyl
cellulose column at pH 7.0 ?
(5 Points)

b) In what order would the oligopeptides Glu-Ile-Pro, Lys-Asp-Phe-Gly, and Val-Trp be eluted from a
gel-filtration chromatography column at pH 7.0 ?
(5 Points)

c) What is the concentration of a lactic acid buffer (pKa = 3.9) that contains 0.25M CH3CH(OH)COOH
and 0.15M CH3CH(OH)COO-1 ? What is the pH of this buffer?
(10 Points)

Page 4

d) Draw the complete structure of the dipeptide Asp-His at pH 7.0.

(10 Points)

e) For an enzyme-catalyzed reaction, the Km is 1 M. When substrate concentration is 100 M, the

initial velocity is 0.1 M min-1. What is the initial velocity when [S] = 2 M?
(10 Points)

Page 5

f) Determine the primary structure (sequence) of a pentapeptide given the following observations:
(10 Points)
(i) amino acid analysis indicated the presence of Q,L,F and Y, in a 2:1:1:1 molar ratio
(ii) Treatment of the peptide with 1-fluro-2,4-dinotrobenzene gave the 2,4-dinitrophenyl derivative
of tyrosine.
(iii) Proteolysis of the peptide with pepsin gave a dipeptide containing Phe and Leu, plus a
tripeptide containing Tyr and Gln.

g) Complete the following table about reversible inhibitors of enzyme-catalyzed reactions: (10 Points)

Inhibitor Type

Apparent Vmax

Apparent KM

Inhibitor binds to E or ES

Competitive

Vmax

KM

Binds to E
Does not bind to ES

Mixed
Uncompetitive

Page 6

(10 Points)
h) For each of the following biochemical reactions, identify the major class of the enzyme catalyzing the
reaction:
I.

II.

glucose-1-phosphate glucose-6-phosphate

Ala-Ser + H2O Ala + Ser

III.

Pyruvate + ATP + CO2 Oxaloacetate + ADP + Pi

IV.

Fructose-1,6-bisphosphate Dihydroxyacetone phosphate + glyceraldehydes-3phosphate

V.

Pyruvate + glutamate -ketoglutarate + alanine

Page 7

(20 Points)
3. For the Citric Acid Cycle, show the part of the pathway that catalyzes the conversion of the
intermediate fumarate to oxaloacetate. For this partial pathway, be sure to include the following
information:
a) names and structures of all intermediates
b) cofactors involved for each reaction.
c) names of all enzymes
d) class of each enzyme

Page 8

(15 Points)
4. The Pyruvate Dehydrogenase complex consists of three different enzymatic components. Complete
the table below with the missing information about the pyruvate dehydrogenase complex.

Enzyme

Enzyme Name

Prosthetic
Group

Additional
Cofactor
(if any)

Reaction Catalyzed

E1

E2

E3

Page 9

(15 Points)
5. For the following reaction proceeding in the forward direction, determine which of the redox couples
is the electron acceptor and which is the electron donor under standard conditions. Calculate the value
of Eo. Determine the free energy change for the reaction.
FAD + 2 cyt c (Fe+2) + 2H+ FADH2 + 2 cyt c (Fe+3)
some physical constants (use only what you need):
Gas constant: R = 1.987 cal/mol K = 8.315 J/mol K
Faraday Constant: F = 96,480 J/V mol
Planks constant: h = 1.584 x 10-34 cal s
Energy conversion factor: 1 cal = 4.184 J

Page 10

(15 Points)
6. Answer each of the following questions (Organize your answers in the following table)
a) List all enzymes of the glycolysis pathway that are not a part of gluconeogenesis.
b) For each of these steps, list the bypass enzyme(s) required during gluconeogenesis starting with
pyruvate. (Reactions or Structures are not required).
c) Identify two enzymes of the gluconeogenesis pathway that are highly regulated.

Glycolysis Enzyme

Bypass Enzyme for

Gluconeogenesis

Is Enzyme Highly Regulated

State Yes or NO

Page 11

(20 Points)
7. A recently discovered bacterium carries out ATP synthesis coupled to the flow of electrons through a
chain of carriers to some electron acceptor. The components of its electron transfer chain differ from
those found in mitochondria; they are listed below with their standard reduction potentials.
Electron carriers in the newly discovered bacterium:

Electrons
E'
Oxidant
Reductant
transferred
(V)

+
NAD
NADH
2
0.32
flavoprotein b (FPb) (oxidized) flavoprotein b (reduced)
2
0.62
3+
2+
cytochrome c (Fe )
1
+0.22
cytochrome c (Fe )
Fe-S protein (oxidized)
Fe-S protein (reduced)
2
+0.89
flavoprotein a (FPa) (oxidized) flavoprotein a (reduced)
2
+0.77

(a) Place the electron carriers in the order in which they are most likely to act in carrying electrons.
(b) Is it likely that O2 (for which E' = 0.82 V) is the final electron acceptor in this organism? Why or
why not?
(c) Calculate the maximum number of ATP molecules that could theoretically be synthesized, under
standard conditions, per pair of electrons transfered through this chain of carriers? (The Faraday
constant, , is 96.48 kJ/V mol; G' for ATP synthesis is +30.5 kJ/mol.)

Page 12

(15 Points)
9. List five high energy bonds (Go of hydrolysis is more negative than -25 kJ/mol). For each of these,
provide the general structure, and an example of a metabolite containing that bond. (Organize your
answer in the following table)

Name of High Energy Bond

General Structure of this Bond

Example of Metabolite
Containing this Bond

Page 13

10. Fill in the blanks for all of the following:

(10 Points)

1. Another name for the citric acid cycle is the _____________________________________ .

2. The pyruvate translocase antiporter allows entry of a molecule of pyuvate into the mitochondrial
matrix, in exchange for a ______________________.

3. In eukaryotes, the enzymes of the citric acid cycle are located in the ____________________.

4. The only membrane-bound enzyme of the citric acid cycle is _________________________.

5. The mechanism of oxidative phosphorylation can be explained by the _________________ theory.

6. Thiamine pyrophosphate is a coenzyme for the ____________________________ enzyme of the

fermentation reaction

7. _____________________ is an inhibitor of ATP synthase.

8. ____________________ accepts electrons from both complex I and complex II.

9. There is no gluconeogenesis in the muscle because muscle cells lack the enzyme ______________.

10. In the multi-subunit ATP synthase molecule, each ________________ subunit(s) has a catalytic site
for ATP synthesis.

Page 14

(15 Points)
11. For EACH of the following, state whether the statement is True or False.
Explain your answer in 1-2 sentences.
a) Oxidative phosphorylation is not possible in the presence of Rotenone, an inhibitor of ETCs
complex I.

b) Glycolysis cannot continue under anaerobic conditions.

c) Mammals cannot convert fatty acids to glucose.

Page 15

* FAD + 2H+ + 2e FADH2

-0.219

***********************************************************************************
Thank you for your co-operation throughout this semester. It was a pleasure teaching you !
I wish you all the best in your careers !
Page 16