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MCB2210 MAY SESSION

1. The resolution limit of a typical fluorescent microscope is ~200 nm. If two ribosomes
that are 50 nm in diameter and 500 nm apart are labeled with a fluorescent antibody, what
will you see?
a. You wont be able to detect the ribosomes at all because they are below the
resolution limit of the microscope.
b. You wont be able to detect them because you need an electron microscope to
see such small objects.
c. You will be able to detect the ribosomes, but they will appear as a single 200
nm spot.
d. You wont be able to detect them because the wavelength of light is not short
enough to resolve them.
e. None of the above.
2. Which would be the best way to visualize dynamics of an intracellular protein in a
living cell?
a. using a fluorescently labeled antibody raised against the protein.
b. expressing a fusion of the protein to GFP
c. using a gold-labeled antibody in combination with transmission electron
microscopy
d. immunoprecipitating the protein from a cytosolic extract
e. none would be useful
3. Antibodies can be used to
a. visualize proteins in fixed and permeabilized cells
b. identify specific proteins on an immunoblot (western blot)
c. precipitate proteins from cellular extracts
d. identify and isolate organelle fractions
e. all are correct.
4. Which is NOT true of phospholipids in a bilayer?
a. Phospholipids are uniform in tail length, making the bilayer a uniform
thickness.
b. Phospholipids can rotate rapidly around their long axis.
c. Phospholipids can easily move from one leaflet to the other leaflet by diffusion.
d. a and b are not true.
e. a and c are not true.

5. Which of the following statements about lipid rafts is false?


a. They can be regions enriched in lipids with saturated fatty acid chains.
b. Because of the different lipid properties, lipid raft formation is an energetically
favorable situation.
c. They can have a different complement of membrane proteins than other regions
of the
bilayer.
d. Lipid rafts can be cholesterol rich regions.
e. All are correct.
6. Which statement(s) about the non-uniformity of lipid bilayers is/are correct?
a. All membranes in a particular eukaryotic cell type have the same composition.
b. A given leaflet of the bilayer of a plasma membrane can vary in its composition
from
location to location.
c. The two leaflets of the lipid bilayer making up the plasma membrane do not
have identical composition.
d. b and c are correct
e. None is correct.
7. You are examining the subcellular localization of a putative membrane protein using
immunofluorescence microscopy. Which is a true statement about what you might
expect?
a. If the antibody you are using was raised to a portion of the protein that is
expected to be exposed to the cytoplasm, you will not need to permeabilize the
membrane with detergent.
b. Although the lipid bilayer is ~ 4nm across, you would expect to observe an
apparent structure ~ 200 nm across due to the resolution limit of light microscopy.
c. If the proteins are located 50 nm apart from each other, you will observe a
dotted appearance of the membrane due to the separation of each labeled protein.
d. None are true.
8. You determine the amino acid sequence of a novel protein. You find that it has a single
alpha helical domain. The alpha helix has primarily hydrophobic amino acids on one face
of the helix, and primarily polar (hydrophilic) amino acids on the other (not the top and
bottom, the sides). Which statement(s) is/ are false?
a. This protein cannot be a single pass transmembrane protein.
b. This protein cannot be a membrane protein.
c. This protein is almost certainly single pass transmembrane protein.
d. This protein could form a multimeric protein complex that spans the bilayer to
allow transport of hydrophilic molecules through the lipid bilayer.
e. b and c are false statements.

9. You engineered a new gene which includes GFP fused to the cytoplasmic domain of a
transmembrane protein. You then added a cardiac-specific promoter and incorporate this
new gene into the genome of the mouse. When you examine cells from these mice in the
fluorescent microscope:
a. You will see the fluorescence throughout the cytoplasm of all the cells of the
mouse.
b. You will see the fluorescence throughout the cytoplasm of all cardiac cells in the
mouse.
c. You will see the fluorescence from the protein in the membrane of all cardiac cells
in the mouse.
d. You will see the fluorescence in the membranes of all the cells of the mouse.
e. All of the above will be seen.
10. You express a recombinant membrane protein and put it into an artificial lipid bilayer.
You tag it with a fluorescent antibody and measure its mobility using FRAP. You find
that it is highly mobile. However, when you do the same experiment on a cell that
naturally expresses that protein, you find that it is not mobile. Which explanation could
account for these results?
a. In the cell, the protein binds to the cytoskeleton.
b. In the cell, the protein binds to other immobile membrane proteins.
c. In the cell, the protein binds to other mobile membrane proteins.
d. a and b could.
e. a and c could.
11. You are studying the cellular basis of protein synthesis. You have two proteinsprotein A and protein B- and you know the following about them. Protein A adopts
different conformations if it is translated in a test tube with purified ribosomes, mRNA,
tRNAs, and amino acids than if it is translated in vivo. When protein B is immunoisolated from cells and denatured by heating in a test-tube, it spontaneously refolds into
the right conformation. Which of the statements below is correct?
a. Protein A but not protein B likely folds with the aid of chaperones and
chaperonins in cells.
b. Protein B but not protein A likely folds with the aid of chaperones and
chaperonins in cells.
c. Both proteins require chaperones and chaperonins to fold properly.
d. Neither protein appears to require chaperones and chaperonins to fold properly.


12. With regard to the diagram above, which statement(s) is/ are correct?
a. Interaction (i) has the smallest Kd.
b. Interaction (i) has the highest affinity.
c. Interaction (iii) has the lowest affinity.
d. Interaction (iii) has the greatest Kd.
e. None are correct.
13. A membrane-associated protein is isolated and found to contain multiple alpha
helices and a large number of hydrophobic amino acids. This protein is most likely a:
a. Transmembrane protein that spans the membrane once.
b. Peripheral membrane protein.
c. Lipid-anchored protein.
d. Transmembrane protein that spans the membrane more than once.
e. None of the above.
14. The permeability to ______ is the most important for generating the peak positive
potential during an action potential.
a. Sodium ions
b. Calcium ions
c. Chloride ions
d. All of the above
e. None of the above
15. Of the steps of the action potential listed, which is the fourth after a cell is stimulated
by a current-passing electrode?
a. Voltage gated sodium channels open.
b. The membrane becomes repolarized.
c. The membrane becomes depolarized to near the sodium equilibrium potential.
d. Voltage gated sodium channels inactivate.
e. Voltage gated K+ channels open.

16. Substance X is an uncharged polar molecule. It is found at higher concentrations


inside of cells than outside. How might substance X enter cells?
a. Diffusion through a channel.
b. Diffusion directly through a lipid bilayer.
c. Active transport.
d. b and c are correct.
e. None of the above.

17. Increasing the activity of vesicular H+-ATPases would:


a. Increase pH inside of vesicles.
b. Decrease pH inside of vesicles.
c. Increase the pH of the cytoplasm.
d. Decrease the pH of the cytoplasm.
e. b and c
18. A given protein is regulated by a protein kinase and a protein phosphatase, and is
active when phosphorylated. Which would you expect to INCREASE the proteins
activity?
a. Expression of a mutant nonfunctional, dominant-negative kinase that blocks the
activity of the cells normal kinases
b. Expression of a mutant nonfunctional, dominant-negative phosphatase that
blocks the activity of the cells normal phosphatases
c. Expression of a mutant hyperactive phosphatase.
d. a and b are correct.
e. b and c are correct.
19. Ras is a small G protein that regulates cell proliferation in response to growth factors.
When Ras is in its active form, cells proliferate. The activity of Ras is carefully regulated
by two other proteins: Ras GEF (guanine nucleotide exchange factor) which stimulates
binding of GTP by Ras, and Ras GAP (GTPase activating protein) which stimulates GTP
hydrolysis by Ras. The activities of these regulatory proteins are, in turn, also regulated.
Ras activity stimulates cell proliferation. Which of the following changes in GAP and
GEF proteins might cause a cell to increase proliferation?
a. A nonfunctional GAP.
b. A permanently active GAP.
c. A nonfunctional GEF.
d. A permanently active GEF.
e. a and d are true.

20. Place the following events in the proper order.


1 Cyclic AMP activates one or more cellular signaling proteins.
2 G-subunit with its attached GTP activates the effector adenylyl cyclase.
3 - Conformational change in the G subunit causing a decreased affinity for the G
subunit.
4 Production of a second messenger, like cAMP.
5 - Replacement of GDP by GTP on the G after interaction with an activated GPCR.
6 - Dissociation of G from the G protein complex.
a.
b.
c.
d.
e.

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21. Ran's role in regulating nucleocytoplasmic transport is based on a mechanism in


which the cell maintains a ______ nuclear concentration and a very low cytoplasmic
concentration of
______.
a. low Ran-GDP, Ran-GTP
b. high Ran-GDP, Ran-GTP
c. high Ran-GTP, Ran-GTP
d. a and c

22. What would you expect to happen if you massively overexpressed a very active
RanGEF in the cytoplasm, such that the normal activity of RanGAP was completely
overridden?
a. Ran would not be recycled by Ntf2
b. Proteins destined for export to the cytoplasm would probably not function
properly, as the cargo/exportin/RanGTP complex would not dissociate.
c. Import of proteins would be accelerated.
d. The importin returning to the cytoplasm would never bind cargo proteins
because it would remain bound to the excess RanGTP present there.
e. a, b and d are correct.
23. Of these five steps listed, what is the FOURTH step in making a nuclear protein?
a. An importin binds to the NLS and moves the protein through the nuclear pore
into the nucleus.
b. The mRNA encoding for the nuclear protein is transported through the nuclear
pore to the cytoplasm.
c. The mRNA is translated.
d. The protein folds.
e. The importin/protein complex encounters Ran-GTP and falls apart.

24. Competition experiments have been used to determine that there are different kinds of
exportins for each species of RNA. You are studying importins for proteins. You
translate in vitro two different nls-containing proteins. Protein A is fused to GFP, while
the other (protein B) is left unlabeled. Which statement below correctly describes results
and interpretations that you might obtain from observing the behavior of these proteins
injected into the cytoplasm of oocytes?
a. Injecting an excess of protein B would not affect import of protein A into the
nucleus if both use the same importin.
b. Injecting an excess of protein B would decrease import of Protein A into the
nucleus if both use the same importin.
c. Injecting an excess of Protein B would not alter import of Protein A into the
nucleus if both use different importins.
d. a and b.
e. b and c.
25. If a nuclear localization signal were added to the gene encoding a lysosmal protein,
where would the protein likely be found?
a. Nucleus
b. Cytoplasm
c. Mitochondria
d. Lysosomes
e. Secreted from the cell.
26. What effect does the docking of the SRP/ribosome/growing polypeptide chain
complex to the SRP receptor have on protein synthesis?
a. The SRP will release from the ribosome.
b. Protein synthesis ceases temporarily.
c. Protein synthesis resumes.
d. The ribosome releases the growing polypeptide chain.
e. a and c are correct.
27. If free ribosomes in the presence of RER vesicles with a mutant, nonfunctional SRP
receptor that cannot bind the SRP, are placed in a test tube with mRNA for a protein with
an n-terminal signal sequence, and everything else needed synthesize proteins, along with
the addition of SRP, where are the proteins found after their production?
a. There are no full-length proteins made.
b. Inside the RER-derived vesicles.
c. Floating free in the aqueous solution in the test tube.
d. Inside microsomes.
e. None of the above.

28. You isolated cellular components to study the transport of proteins in a cell. After a
lot of hard work, you take a vacation, but you forget to write down how far along in the
purification you progressed. If you put some of your preparation in a test tube, and then
add mRNA for a secretory protein, you find that the mRNA is completely translated and
the new protein is sensitive to protease. Your preparation has which of the following?
a. Ribosomes, SRP, microsomes, and SRP receptor.
b. Ribosomes, SRP, and microsomes, but no SRP receptor.
c. Ribosomes and microsomes, but no SRP.
d. Ribosomes, SRP, but no microsomes and no SRP receptor.
e. SRP and microsomes with SRP receptor, but no ribosomes.
29. Which of the following could prevent a secretory protein from being secreted
normally?
a. A mutation that causes addition of mannose-6-phosphate to the oligosaccharide
structure.
b. A mutation that prevents addition of N-linked oligosaccharides in the ER.
c. A mutation that results in deletion of the start transfer sequence.
d. Addition of an ER-retrieval sequence.
e. All of the above.
30. Which of the following are made on RER-bound ribosomes?
a.Nuclear proteins
b.mitochondrial proteins
c.soluble proteins found inside lysosomes
d.secretory proteins
e. c and d
31. Referring to the picture below showing abnormal final localizations of a normally
secreted protein, loss of function mutations in which molecule(s) could result in
phenotype 1?
a. SRP.
b. Translocation channel.
c. Clathrin proteins.
d. Mutations in a and b could result in phenotype 1.
e. All of the above could result in phenotype 1.

32. The following are involved with synthesis of a secreted protein. Which is the
FOURTH step of the five listed?
a. The SRP dissociates from the ribosome and start transfer sequence
b. The SRP binds to the start transfer sequence.
c. The signal (start transfer) sequence is removed by signal peptidase.
d. The SRP binds to SRP receptor
e. Protein synthesis halts temporarily
33. If you engineered a new protein by taking the gene for a single pass transmembrane
protein that has an n-terminal signal sequence and remove the stop transfer sequence
from the middle of the gene, what is a possible destination of the new protein?
a. It would be found in the cytoplasm.
b. It would be glycosylated and transported to mitochondria.
c. It would be found in the nucleus
d. It would be found as a single pass membrane protein in the plasma membrane.
e. None is a possible destination.
34. If a nuclear localization signal were added to the gene encoding a plasma membrane
protein, where would the protein likely be found?
a. Nucleus.
b. Plasma membrane.
c. Cytoplasm.
d. Lysosomes.
e. Secreted from the cell.
35. What does UGGT do if it binds to a misfolded glycoprotein?
a. It interacts with Calnexin and ERp57 and transports the protein to the proteasome.
b. It adds a single mannose back to one of the glucose residues at the exposed end of
the recently trimmed oligosaccharide.
c. It adds a single glucose back to one end of the recently trimmed oligosaccharide.
d. It degrades the protein.
e. a and b are correct.

36. How are proteins imported into mitochondria and chloroplasts?


a. Pre-translationally.
b. Imported fully-formed and properly-folded across membrane of the appropriate
organelle.
c. Post-translationally.
d. Post-transcriptionally.
e. b and c are correct.

37. Put the following in the correct order:


I. Protein is imported through the TIM complex
II. Protein is imported through the TOM complex
III. Fully-folded, mature protein is located in the mitochondrial matrix
IV. Protein is folded by mitochondrial chaperone using ATP
V. Import signal is recognized by receptor protein
a. V, I, II, IV, III
b. II, V, III, I, IV
c. II, III, V, IV, I
d. V, II, I, IV, III
e. none of the above is correct.
38. If you took a gene that is normally present in the mitochondrial genome and codes for
an enzyme that works in the matrix, and then inserted the gene into the nucleus of a cell,
what would you expect the final destination of the protein to be?
a. ER lumen
b. Nucleus.
c. Secreted.
d. Mitochondrial matrix
e. None of the above
39. If a patient has a lysosomal storage disease in which all lysosomal proteins are found
secreted from the cell, and those proteins have no Man-6-phosphate attached to their
oligosaccharides, then the defect is most likely:
a. In the SRP receptor.
b. In the enzyme that adds man-6-p to lysosomal proteins.
c. In the man-6-p receptor
d. In the lysosomal enzyme RNAase.
e. In the enzyme that co-translationally glycosylates proteins.
40. Which is the FOURTH step of the five listed in lysosomal enzyme sorting?
a. Co-translational addition of an oligosaccharide to the protein.
b. Binding of Man-6-Phosphate to its receptor in the TGN.
c. Removal of the man-6-phosphate signal
d. Transport of the enzyme from the TGN
e. Transport from the ER to the Golgi
41. PTS1- and PTS2-bearing matrix proteins target to
a. a common cytosolic receptor.
b. a common translocation machinery on the peroxisomal membrane.
c. a common receptor on the nuclear pore that catalyzes entry into the nucleus via
pore targeting sequences.
d. a common receptor protein within the peroxisomal matrix that activates protein
processing for PTS1- and PTS2-bearing proteins.

42. Which of the models below suggests that the Golgi cisternae are transient structures
that form at the cis face and travel through the Golgi complex, moving physically and
exiting the organelle at the trans face, while changing during the journey?
a. the cisternal maturation model
b. the cargo carrying model
c. the vesicular transport model
d. the secretory transport model
e. the chemiosmotic model
43. What is the function of the protein coat on budding vesicles?
a. It acts as a mechanical device that helps to form the vesicle.
b. It protects the forming vesicle from osmotic shock.
c. It prevents the diffusion of harmful materials into the forming vesicle.
d. It provides a mechanism for selecting components and cargo to be carried by
each vesicle.
e. a and d
44. Which coated vesicles are NOT involved in moving materials in a retrograde
direction from the Golgi complex to the ER?
a. COPII-coated vesicles
b. COPI-coated vesicles
c. clathrin-coated vesicles
d. a and b
e. a and c
45. Which of the following polypeptides would be expected to be glycosylated?
a. A histone protein bound to nuclear DNA.
b. A lysosomal enzyme.
c. COPI vesicle coat protein.
d. The SRP.
e. A cytoplasmic chaperone.
46. What do you think will be the effect on COPI-coated vesicles if a cell is treated with
GTP analogues that can bind to the G-protein involved in vesicle formation, but cannot
trigger its active conformation?
a. They will accumulate in the nucleus.
b. They will accumulate in the cytoplasm.
c. They will fuse into one giant vesicle that was seen in the cytoplasm.
d. They will decrease substantially in number in the nucleus.
e. They will decrease substantially in number in the cytoplasm.

47. All of the following could prevent a secretory protein from being secreted normally
EXCEPT:
a. A mutation that results in the deletion of the start transfer sequence.
b. The addition of an ER-retrieval sequence.
c. A mutation that causes addition of mannose-6-phosphate to the carbohydrate
tree structure.
d. A mutation that prevents addition of N-linked oligosaccharides in the ER.
e. Glycosylation of the protein in the ER.
48. What likely determines the specificity of vesicle fusion to a target membrane?
a) interactions between tethering proteins alone
b) interactions between Rabs alone
c) interactions between specific combinations of interacting proteins, including
tethering proteins, Rabs and SNAREs
d) interactions between SNAREs alone
e) a single protein in the membrane of the particular vesicle and target membrane

49. Overall, cell surface area would DECREASE if:


a. The rate of clathrin-coated pit formation increased
b. The rate of clathrin coated pit formation decreased
c. The rate of constitutive exocytosis increased
d. The rate of constitutive exocytosis decreased
e. a and d are true
50. You are tracing the path of the LDL receptor for cholesterol transport from its
synthesis to its final destination on the plasma membrane. The order of listed
compartments in which you would find it is:
I. medial Golgi
II. Trans Golgi network
III. Secretory vesicles
IV. cis-Golgi
V. Rough ER
a. V, IV, II, I, III
b. II,V, IV, I, III
c. V, IV, I, II, III
d. V, I, IV, II, III
e. V, II, I, IV, III