Iram, S.H. and Cole, S.P.C. 2011. Expression and Function of Human MRP1 (ABCC1) Is Dependent on Amino Acids in Cytoplasmic Loop 5 and Its Interface with Nucleotide Binding Domain 2. J. Biol. Chem. 286: 7202-7213.
Using the values for ?G’ in each case, calculate the equilibrium constants K’eq for the following reactions which occurred at pH 7.0 and 25oC. (6 marks)
ATP + H2O doublearrow_image AMP + PPi ?G’ = -45.6 kJ/mol
malate doublearrow_image fumarate + H2O ?G’ = +3.1 kJ/mol
Draw the following amino acids linked by peptide bonds: (4 marks)
Aspartate
Leucine
Cysteine
Threonine
The amino acid sequence shown below represents a portion of a peptide obtained from a large protein.
Leu(1) His(2) Ile(3) Thr(4) Arg(5) Phe(6) Phe(7) Pro(8) Cys(9) Met(10) Gly(11) Glu(12) Ala(13) Ile(14) Pro(15) His(16) Thr(17) Glu(18) Asp(19) Cys(20) Gln(21) Met(22) Ile(23) His(24) Cys(25) Pro(26) Arg(27) Lys(28) Gln(29) Pro(30) Tyr(31) Leu(32)
Determine where the chain would be more likely to show ß bends or reverse turns. Provide a clear explanation for your answer. (5 marks)
Discuss where one would find cross-linked disulfide bonds in this sequence.
(3 marks)
Would you consider this molecule to be in an a-helix formation? Explain your answer.(2 marks)
List 5 roles for proteins in vivo and give and specific example of a protein for each.
(5 marks)
Match each of the descriptive phrases in Column B with the appropriate term in Column A. (10 marks)
a. Peptidoglycan ______ glucose with a(1 ? 4) and a(1 ? 6) bonds found in plants
b. Lactose ______ highly branched polymer of linear glucose molecules
c. Anomers ______ glucose units, H-bonds for strength
d. Amylopectin ______ main component of a bacterial cell wall
e. Glycogen ______ is a heteropolymer composed of D- glucoronate and N-acetyl-D-glucosamine that acts as shock absorber and lubricant.
f. Chitin ______ carbonyl group on monosaccharide is an aldehyde
g. Hyaluronic acid ______ a-D-galactose and ß-D-galactose
h. Aldose ______ disaccharide in dairy products
i. Cellulose ______ carbonyl group on monosaccharide is a ketone
j. Ketose ______ N-acetyl-D-glucosamine units, H-bonds
What are bacterial cell walls composed of? Describe the biochemical structure. How does the antibiotic penicillin act to disrupt bacterial cell walls? (5 marks)
Distinguish between a micelle, a bilayer, and a liposome. What are the forces that maintain these structures? (4 marks)
Describe the structure of a biological membrane. What are the types of proteins found associated with the membrane? Describe the mobility of proteins and lipids in the membrane. (5 marks)
How is the fluidity of a biological membrane maintained? What is the function of cholesterol in the membrane? (4 marks)
Outline the mechanism of pumping Na+ and K+ across the cell membrane. (4 marks)
Draw the structures of all of the nitrogenous bases found in DNA and RNA. (5 marks)
If a DNA molecule comprised the following sequence, what would be the complementary sequence? If the DNA sequence was transcribed to RNA, what would be the mRNA sequence? (2 marks)
5’-ATGCAACGCTTCAAGATAGGCATCTAA-3’
What is a plasmid? Describe how plasmids are used in genetic engineering. (3 marks)
An experiment was performed to determine the effects of an inhibitor on the breakdown of glycogen by an enzyme. In an accompanying experiment, the inhibitor was added to the glycogen-enzyme suspension and reacted using the same experimental conditions. The data obtained from these experiments is tabulated below.
Glycogen (mM) Product Formed Glycogen only (mM/min) Product Formed Glycogen and Inhibitor (mM/min)
0.5 22.6 15.9
1.0 32.1 24.9
1.5 38.1 28.2
2.0 41.2 35.1
2.5 44.7 40.0
3.0 48.5 43.3
Draw the Michaelis-Menten and Lineweaver-Burke plots of these data. (6 marks)
Determine the form of inhibition observed from these results and explain your rationale for this form. (3 marks)
Determine the values for Km and Vmax from these results.
(5 marks)
To answer the following questions, read the paper by Iram and Cole in the Digital Reading Room.
What hypothesis do the authors propose? (2 marks)
Describe the structure and outline the function of MRP1. (3 marks)
What is the CL5 region? For the purposes of this study how did the authors define the CL5 region? What amino acids comprise this region? (4 marks)
Briefly, how did the authors determine the role of the CL5 region in the expression and activity of MRP1? (6 marks)
What were the overall results of their findings? (2 marks)
What is the implication of this study? (2 marks)
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