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BISC 3754 Lecture Notes - Lecture 12: Secretion, Copii, Triskelion

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yellowweasel253
28 Apr 2018
School
Fordham University
Department
Biological Sciences
Course
BISC 3754
Professor
Zencheck

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1
Intracellular Vesicular Traffic I
● Lumen of vesicle is the same as the extracellular region
Exocytosis- transport vesicle fuses with plasma membrane, contents released into EC space
Endocytosis- plasma membrane internalizes, creates transport vesicle
Secretory pathway: rough ER → golgi → secretory vesicles → extracellular space
→ early endosome → extracellular space
→ late endosome → lysosome
Endocytic pathway- Plasma membrane to early endosome to late endosome to lysosome
● Molecular markers on cytosolic surface guide traffic
○ Allow transport vesicles to correctly fuse
○ Donor compartment membrane protein orientation is maintained
● Transport vesicles bud as coated= distinct protein coat on cytosolic side
○ Lost after it pinches off
○ Functions:
■ Inner coat layer selects appropriate mem molecules for transport
■ Outer coat layer shapes vesicle
● Clathrin coated- transport from golgi to plasma mem, between Golgi and endosome
● COPI/COPII- transport from ER to golgi
○ COPI- bud from golgi
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○ COPII- bud form ER
● Vesicles need a coat protein
○ Has protein tag (recognition)
○ Cargo has protein binding sites/receptors
○ GTP binding protein- monomeric GTPase (small G protein)
Clathrin
● 3 large and 3 small polypeptide chains in each subunit= triskelion framework
● Adaptor proteins bind clathrin coat to membrane and trap proteins (between cage and mem.)
○ Adaptors are specific for cargo receptors
AP2 adaptor protein
● Each AP2 has 4 subunits and can bind to 4 PIP2
● Phosphoinositide P1(4,5)P2 in membrane (phospholipid)
○ AP2 binds to P1(4,5)P2→ changes conformation= binding sites for receptors exposed
● Cargo receptors also in membrane
○ Binding signals endocytosis
○ Cooperative process= more AP2 bind and coats are assembled once it starts
■ Due to curvature forming in membrane
● Phosphoinositides (PIP) → rapid phosphorylation/dephosphorylation
○ Distribution varies among organelles
○ PIP binding proteins regular vesicle formation and traffic
○ Reactions occur a 3, 4, or 5 carbon of sugar (inositol) head
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Document Summary

Lumen of vesicle is the same as the extracellular region. Exocytosis- transport vesicle fuses with plasma membrane, contents released into ec space. Secretory pathway: rough er golgi secretory vesicles extracellular space. Endocytic pathway- plasma membrane to early endosome to late endosome to lysosome. Molecular markers on cytosolic surface guide traffic. Donor compartment membrane protein orientation is maintained. Transport vesicles bud as coated= distinct protein coat on cytosolic side. Inner coat layer selects appropriate mem molecules for transport. Clathrin coated- transport from golgi to plasma mem, between golgi and endosome. Gtp binding protein- monomeric gtpase (small g protein) 3 large and 3 small polypeptide chains in each subunit= triskelion framework. Adaptor proteins bind clathrin coat to membrane and trap proteins (between cage and mem. ) Each ap2 has 4 subunits and can bind to 4 pip2. Ap2 binds to p1(4,5)p2 changes conformation= binding sites for receptors exposed. Cooperative process= more ap2 bind and coats are assembled once it starts.

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Related Questions

1/What is the fate of a protein with no N-terminus Signal Sequence?

(a)Transport through the constitutive secretory pathway

(b)Dispersal throughout the cytoplasm

(c)Transport into the nucleus

(d)Transport to the rough endoplasmic reticulum (RER)

2/ In nerve cells, Botulinum toxin-A degrades a subunit of a T-snare localized on the plasma membrane, how would this affect vesicular trafficking?

(a) It blocks the co-translational import of neurotransmitter proteins into the Endoplasmic reticulum via the translocon.

(b) It prevents the addition of carbohydrates to translated proteins

(c)It blocks the fusion of neurotransmitter-containing vesicles with the plasma membrane because theT-snare would not be able tophysically interact with the V-snare.

(d) None of the above

3/

What is the correct order of events of Clathrin-mediated endocytosis?

1. Uncoating of Clathrin- coated vesicle

2. Recruitment of Clathrin and selection of cargo

3. Fusion of vesicle with early endosome

(a) 1, 2, 3

(b) 2, 1, 3

(c) 2, 3, 1

(d) 3, 1, 2

4/ Vesicles traveling anterogradely will eventually fuse with the cell membrane.

Which of the following elements does NOT participate in the fusion event?

(a) v-snare

(b) t-snare

(c) SNARE complex

(d) clathrin coat protein

5/

When a cell releases a hormone into the bloodstream and a number of cells in a

distal location respond, this type of signaling is...

(a) Autocrine signaling

(b) Endocrine signaling

(c) Paracrine signaling

(d) Juxtacrine signaling


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

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.
(
)
(