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Biochemistry 3381A Lecture Notes - Lecture 5: Signal Transducing Adaptor Protein, Clathrin, Protein Structure

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spectral
20 May 2020
School
Western University
Department
Biochemistry
Course
Biochemistry 3381A
Professor
Brian Shilton

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Document Summary

Clathrin binds: large part controlled by lipid binding, ap2 in the locked position contains 2 and 2 in the horizontal position, phosphoinositide and cargo receptors are located on the plasma membrane itself. If signal sequence is mutated, receptor doesn"t go inside the cell. Amino acid sequences signal sequences or signal patches. Entire endocytosis process takes around a minute to happen. Pinching off of the plasma membrane: pinching of vesicle is enabled by another protein dynamin (gtpase). Experiment done on fruit flies through wildtype and mutated type, and heated up to a certain temperature. Almost all flies with the mutation were paralysed but recovered once temperature decreased again. Copii coat is less dense than clathrin: clathrin used in endocytosis, need to bend plasma membrane which is quite rigid, secretory pathway more flexible membranes, so copii coat is less dense than clathrin coat.

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Related Questions


1. Name two similarities and two differences between thecellular processes of importing

protein into the ER and importing protein to the nucleus.

2. What proteins are directly involved in the transportation ofcargo in a clathrin-coated

vesicle?

a. Dynamin

b. Adaptins

c. Cargo receptors

d. t SNARES

3. Suppose a cell was treated with colchiine, a drug causingmicrotubule disassembly.

What would happen within the cell? Why?

Suppose there were a genetic defect in the genes that code forkinesins and dyneins.

What would happen within the cell? Why?

4. Which of the following are true about microtubules?

a. They are utilized by the disease causing bacteriaLysteria.

b. Instability in the minus-end of the microtubule is caused bythe hydrolization of GTP.

c. Centrosomes contain binding sites for microtubules composed atype of tubulin ring.

d. Cellular concentrations of tubulin are above the criticalconcentration for plus-end

polymerization.

e. Motor proteins can attach organelles to the microtubules.

5. Indicate which of the following statements are false:

a. Membrane fusion is energetically unfavorable and thusrequires ATP or similar high

energy molecule to occur

b. Clathrin is responsible for capturing specific molecules fortransport in coated vesicle

structures.

c. Receptor proteins that are incorporated into an endosome canbe retrieved back to the

membrane domain it originated from; can return to a new domainof the membrane; or

can enter the lysosome for digestion.

d. Transport vesicles occasionally bud from the trans Golginetwork to fuse with the

plasma membrane in a process called constitutive exocytosis.

e. Most components of the endocytic vesicle membrane eventuallyare returned to the

plasma membrane for reuse.

6. Match the following organelles with their function:

Organelle Bank:

a. Mitochondria

b. Endoplasmic reticulum

c. Nucleus

d. Lysosome

e. Endosome

f. Peroxisome

g. Golgi apparatus

_____ quality control of mRNA

_____ location of oxidative phosphorylation

_____ responsible for detoxifying organic molecules (livercells)

_____ houses and protects genetic material

_____ responsible for modification and sorting of proteins andlipids

_____ location of ATP synthesis

_____ responsible for sorting endocytosed materials

_____ site of degradation and digestion

_____ responsible for oxidizing toxic molecules

_____ location of lipid synthesis

_____ location of hormone synthesis (adrenal cells)

B. Which organelle(s) from the above list communicate(s) withother organelle(s)

through the use of vesicular transport? ________________

C. Which organelle(s) from the above list receive proteins madein the

cytosol?___________________

D. Through which organelle(s) must proteins pass to reach theorganelle(s) which do not

receive proteins made exclusively in thecytosol?______________

E. Entrance into which organelle(s) requires that proteinsunfold and snake through the

membrane? _________________

F. How do organelles maintain their position in the cytosol? Bespecific.

G. Give 2 advantages that eukaryotic cells gain by havingorganelles.

7. Which of the following statements are true regardingactin?

a. They are arranged in a

The vesicles that transport materials from donor membranes to acceptor membranes include:

1.

COP-II coated vesicles
2.

COP-III coated vesicles
3.

COP-I coated vesicles
4.

Clathrin coated vesicles

2.5 points

QUESTION 11

During skeletal muscle contraction, the roles of ATP include:

1.

ATP is not required for forming the crossbridges between myosin and actin filaments.
2.

ATP is utilized as a structural linker to mediate the interaction between myosin and actin filaments
3.

Hydrolysis of ATP provides energy for myosin head to move along actin filaments and the binding of ATP to myosin head allows the detachment of myosin fromactin filaments.
4.

ATP binds to actin filaments to drive the power stroke of myosin head.

2.5 points

QUESTION 12

One of the functions of smooth endoplasmic reticulum is

1.

Synthesizing steroid hormone in the endocrine cells of the gonad and adrenal cortex
2.

Serves as the starting point of the biosynthetic pathway
3.

Synthesizing secretory proteins
4.

Degrading cell debris

2.5 points

QUESTION 13

The function of COP-II coated vesicles is mainly:

1.

Transporting materials from endosomes to Golgi complex
2.

Moving materials from the ER to the ERGIC and Golgi complex.
3.

Transporting materials from Golgi complex back to endoplasmic reticulum.
4.

Involved in receptor-mediated endocytosis.

2.5 points

QUESTION 14

Which of the following processes is completed in rough endoplasmic reticulum?

1.

Chaperones help misfolded proteins fold correctly.
2.

Proteins that target to nucleus are synthesized in rough endoplasmic reticulum.
3.

O-linked glycosylation occurs in rough endoplasmic reticulum.
4.

Misfolded proteins are destructed in rough endoplamic reticulum.

2.5 points

QUESTION 15

Kinesin is the motor protein that coorporates with microtubules. It has the following functions:

1.

It uses GTP as energy source.
2.

It can hydrolyze ATP and use the energy to drive cargo along microtubules from minus end to plus end.
3.

It has a globular head that can act as GTPase
4.

The tail of kinesin is more diverse and binds to cargo

2.5 points

QUESTION 16

For correct segregation and trafficking, lysosomal proteins are tagged in the cis-Golgi with phosphorylated mannose residues, and then the tagged lysosomal enzymes are recruited to clathrin coated vesicles by mannose 6-phosphate receptors.

True

False

2.5 points

QUESTION 17

The process that specialized cells, such macrophages, engulf relatively large particles is:

1.

Called pinocytosis
2.

Called phagocytosis
3.

Mediated by COP-II coated vesicles
4.

Called autophagy

2.5 points

QUESTION 18

Proteins synthesized on rough endoplasmic reticulum and smooth endoplasmic reticulum are imported co-translationally to their destinations

True

False

2.5 points

QUESTION 19

Treatment of cells with a drug that promotes microtubule disassembly disperses Golgi complex into separate stacks of membrane compartments, suggesting that:

1.

Golgi complex is made of microtubules.
2.

Microtubules are important for the organization of Golgi complex.
3.

Microtubules are transported into Golgi complex
4.

Golgi complex is composed of tubulin proteins

2.5 points

QUESTION 20

Which of the following is correct about signal sequence of a secretory protein?

1.

It is an “address codes” for protein trafficking pathways
2.

It is usually located at the C-terminus.
3.

It targets the ribosome and the protein that is being synthesized to endoplasmic reticulum.
4.

It is removed in Golgi complex.
True or False? True False
1. Proteins rapidly pass artificial membrane (liposome).--------------------------------------------------------------------------------------( ) ( )
2. Amphiphatic detergent such as SDS spontaneously forms bilayer inwater. ------------------------------------------------------------( ) ( )
3. Cholesterol does have a hydrophilic head and hydrophobic tail.-------------------------------------------------------------------------( ) ( )
4. Fat molecules are one of the major components of the plasmamembrane.------------------------------------------------------------- ( ) ()
5. Phospholipid molecules such as phosphatidylcholine spontaneouslyform bilayer. ----------------------------------------------------( ) ( )
6. Flippase facilitates lateral movement of phospholipids withinthe plane of the membrane.----------------------------------------- ( ) ( )
7. Glycolipids make bilayer less permeable to small water solublemolecules.----------------------------------------------------------- ( ) ()
8. Membrane inner leaflet is enriched with phosphatidylcholine,Sphingomyelin, and Glycolipid. -----------------------------------( ) ( )
9. Plasma membrane proteins can have an enzyme activity.--------------------------------------------------------------------------------( ) ( )
10. A single pass membrane protein must have an amphiphatic alphahelix.-------------------------------------------------------------- ( )( )
11. Plasma membrane proteins can be linked to a lipid molecule atthe outer leaflet of the plasma membrane. --------------------- ( )( )
12. The lateral motility of membrane proteins can be restricted bytight junctions.------------------------------------------------------ ( ) ()
13. Channel proteins have specific binding sites for ions and thusslower than carrier proteins.--------------------------------------- ( ) ( )
14. An antiport would function as a symport if its orientation inthe membrane were reversed. ---------------------------------------( ) ( )
15. A transmembrane pore can be formed by multiple amphipathicalpha helices. ----------------------------------------- ( ) ()
16. Porin proteins form water-filled channels in the outer membraneof a bacterium and are mostly composed of beta sheets. ---- ( ) ()
17. Glucose uniporters localize at the apical side of theintestinal epithelium.------------------------------------------------------------ ( ) ()
18. GAP junction is a selective pore for anions.-----------------------------------------------------------------------------------------------( ) ( )
19. A voltage-gated Na+ channel adopts only two conformationstates: open and closed.----------------------------------------------- ( ) ( )
20. Voltage gated K+ channel opens faster than the Voltage gatedNa+ channels.-------------------------------------------------------- ( ) ()
21. Opening of chloride channel at the synapse will help to fire anaction potential.------------------------------------------------------ ( ) ()
22. The nuclear pore complex forms a gate through which moleculespass the nucleus uni-directionally. -------------------- ( ) ()
23. A completely folded protein is imported into a mitochondrionthrough a large non-selective pore.--------------------------------- ( ) ( )
24. ER signal sequence contains multiple lysine and arginineresidues.----------------------------------------------------------------------( ) ( )
25. The plasma membrane is a sorting station of inward endocyticpathway.---------------------------------------------------------------- () ( )
26. The trans Golgi functions as a sorting station of outwardsecretory pathway.----------------------------------------------------------- ( ) ()
27. Vesicles budding from ER are coated with Clathrin.----------------------------------------------------------------------------------------( ) ( )
28. RabGTPases and tethering proteins are the major specificitydeterminant of vesicle transport system. --------------------------( ) ( )
29. v-SNARE proteins are one of the tethering proteins.------------------------------------------------------------------------------------( ) ( )
30. In secretory cells, the constitutive and regulatory pathways ofexocytosis diverge at the cis Golgi network. ------------------ ( )( )
31. Many proteins are posttranslationally glycosylated at tyrosineresidues in the ER.------------------------------------------------- ( ) ( )
32. Cells import LDL molecules exclusively through pinocytosis.-------------------------------------------------------------------------( ) ( )
33. Cells recycle LDL receptors from lysosome to the plasmamembrane.----------------------------------------------------------------- () ( )
34. Phagosomes fuse with late endosome.------------------------------------------------------------------------------------------------------( ) ( )
35. Small vesicles from ER near the plasma membrane can fuse withthe plasma membrane upon bypassing Golgi apparatus. -- ( ) ()
36. If a Sec mutant showed an accumulation of newly synthesizedproteins at ER,
then the Sec gene is likely a component of vesicle-buddingmachinery of trans-Golgi network.------------------------------------ ( ) ( )
37. A signal recognition particle is a single pass trans-membraneprotein of the ER membrane.---------------------------------------- ( ) ( )
38. Typical animals cells display higher levels of intracellularsodium than extracellular compartments.------------------------------ ( ) ( )
39. Typical animals cells display higher levels of intracellularchloride than extracellular compartments.----------------------------- ( ) ( )
40. The Na-K Pump is a GTPase that can adopt active and inactiveconformational states.----------------------------------------------- ( ) ( )
1. Phospholipase
C-­-ß
produces
phosphatidylinositol
3,4,5-­-trisphosphate
and
diacylglycerol.
(
)
(
)
2. PDGF
activates
GPCR.
-­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­----­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­-
(
)
(
)
3. Rho
is
an
example
of
heterotrimeric
G
proteins.
-­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­----­--­--­--­--­--­-
(
)
(
)
4. All
members
of
the
G
protein
coupled
receptors
interact
with
small
GTPase
Ras.
-­--­--­--­--­--­--­--­--­--­-
(
)
(
)
5. Adrenaline
is
an
example
of
the
ligands
for
the
RPTK
family.
-­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­-
(
)
(
)
6. Ras
GEF
activates
RPTK.
-­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­----­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­-
(
)
(
)
7. PH
domain
containing
proteins
bind
to
activated
EGFR.
-­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­-
(
)
(
)
8. Ligands
for
GPCR
proteins
induce
dimerization
of
the
receptor.
-­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­-
(
)
(
)
9. Grb2
is
an
adaptor
protein
that
binds
to
a
phospho-­-tyrosine
residue
of
activated
RPTK
through
a
PH
domain.
-­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­----­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­-
(
)
(
)
10. When
a
cell
is
expressing
a
large
number
of
dominant
negative
EGFR
proteins,
an
introduction
of
a
constitutively
active
Ras
mutant
will
allow
EGF
signaling.
-­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­-
(
)
(
)
11. PI3K
generates
phosphatidylinositol
3,4,5-­-trisphosphate
from
IP3.
-­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­-
(
)
(
)
12. Calcium
inhibits
PKC.
-­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­----­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­-
(
)
(
)
13. cAMP
activates
PKA.
-­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­----­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­-
(
)
(
)
14. The
MAP
kinase
ERK
inhibits
MAP
kinase
kinases.
-­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­----­--­-
(
)
(
)
15. cAMP
is
an
example
of
a
first
messenger.
-­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­----­--­--­--­--­--­--­--­--­--­--­--­--­--­--­-
(
)
(
)
16. All
7-­-Transmembrane
receptors
are
G
Protein
Coupled
Receptors.
-­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­--­-
(
)
(
)
17. Both
Ga
and
Gß
subunits
of
the
heterotrimeric
G
protein
can
mediate
GPCR
signaling.
-­--­--­-
(
)
(
)
18. Both
Ga
and
Gß
subunits
of
the
heterotrimeric
G
protein
have
GTPase
activity.
-­--­--­--­--­--­--­--­--­--­--­--­-
(
)
(
)
19. The
Gas
subunit
of
the
heterotrimeric
G
protein
suppresses
the
function
of
Adenylyl
cylase.
(
)
(