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z:Cell 300 > Lecture 9: Trans-Golgi Network (TGN) > Flashcards

Lecture 9: Trans-Golgi Network (TGN) Flashcards

1
Q

TGN is the most ___

A

distal region of the golgi complex

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2
Q

TGN functions to modify and sort ____ from ____ and proteins & lipids from ____

A

newly synthesized proteins & lipids
-proximal golgi
-endocytic compartments

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3
Q

defects in sorting/release of cargo in TGN associated which diseases?

A

-autoimmune disease, cancer, neuropathology, diabetes

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4
Q

______ of viral proteins affect pathogenesis

A

TGN processing

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5
Q

major function of TGN

A

-sort proteins (localization) but also modifies proteins

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6
Q

TGN is _____ & _____distinct from golgi stack

A

-morphologically & biochemically

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7
Q

morphological differences between TGN and golgi stack

A

-tubular network and associated vesicles
-cisternae are not flattened against golgi stack
-cisternae has peeled away from trans cisternae of golgi stack

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8
Q

biochemical differences between TGN and golgi stack

A

-osmium reduced by unsaturated lipids in cis-golgi (reducing environment)
-TGN has acid phosphatase (acidic environment)

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9
Q

TGN is considered _____ by many reasearchers

A

separate entity

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10
Q

evidence that TGN is separate

A

-TGN reacts differently to drugs and temperatures that impair protein/membrane trafficking

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11
Q

BFA (Brefeldin A) causes ___ (for golgi stack +TGN)

A

-TGN fuses with endosomal membranes
-Golgi stack fuses with ER

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12
Q

Membrane an protein cargo blocked in TGN and ER at which temperatures?

A

-TGN: 20 C
-ERGIC: 15 C

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13
Q

Enzymes of TGN do what?

A

-terminal glycosylation
-sulfation
-endoproteolysis

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14
Q

TGN: terminal glycosylation (def.) + why?

A

-addition of sialic acid to N-linked oligosaccharides by dialytransferase (ST)
-keeps negative charge on cell surface
-keeps water at cell surface

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14
Q

TGN: sulfation (def.)

A

-addition of sulfate groups to Tyrosine
-can affect protein-protein interactions

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15
Q

human brain is rich in ____

A

sialic acid (4X) for neurotransmission + memory

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16
Q

loss of tyrosine sulfation can affect ___

A

blood clotting (VIII)

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17
Q

Furin is ___

A

a transmembrrane protein in TGN that cleaves pro-proteins

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18
Q

Furin cleaves where?

A

R-X-(R/X)-R

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19
Q

Furin + COVID-19

A

-furin cleavage of spike protein promote intereaction of virus particles with receptor ACE2 by exposing receptor binding domain (RBD)

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20
Q

presence of cleavage site associated with _____

A

highly pathogenic influenza virus strains

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21
Q

furin cleaves what?

A

mammalian, viral, bacterial substrates

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22
Q

furin inhibitors are good antivirals but there is a problem: ____

A

-furin does many important function in mammals but viruses take advantage of it

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23
Q

TGN is a _____ site for proteins/lipids in eukaryotic cells

A

major sorting site

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24
Q

3 main sorting pathways of TGN +examples + signal needed?

A

-to endosomes (ex. lysosomal enzymes) + signal needed
-to secretory granules (ex. regulated secretion) + signal needed
-constitutive transport to PM + no signal needed (except in polarized cells)

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25
Q

Alzheimer disease (AD) is caused by ___ due to

A

accumulation of plaque that contain AB oligomers due to abnormal cleavage of amyloid precursor protein by B-secretase (makes AB oligomers)

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26
Q

Production of AB oligomers happens in ___ because ____

A

-TGN
-b-secretase which are normally localized in endosomes but decrease in GGA or increase in retromer make B-secretase localize in TGN

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27
Q

AB plaques make by ___ (2)

A

-B-secretase
-gamma-secretase

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28
Q

If movement of beta-secretase to TGN ___ or b-secretase to endosome is ____, cleavage of APP increases

A

-increases
-blocked

29
Q

normal cleavage of APP is by ___

A

a-secretase

30
Q

live cell imaging allows for ____

A

visualization of cargo and interaction with membrane compartments and cytoskeleton

31
Q

temperature blocks are useful for ___

A

synchronizing cargo flow

32
Q

temperature block of TGN

A

20 C

33
Q

coat protein for constitutive transport from TGN

A

-no known coat protein

34
Q

carriers size for constitutive transport

A

-large structure (170 -1700nm) (half size of cisternia)
-variable size

35
Q

shape of carriers for constitutive transport

A
36
Q

constitutive transport is ____sensitive and regulated by ___

A

-BFA
-ARFs (small GTPases)

37
Q

constitutive transport requires ___motores (____), ____and ___

A

-molecular motors
-kinases
-rabs

38
Q

kinesin help with the movement of ____

A

carriers

39
Q

constitutive transport to PM: sorting signals ___

A

not needed (default pathway)

40
Q

VSG is a ___ targeted protein

A

basolaterally

41
Q

post TGN cargo (constitive transport) travels along ____

A

microtubules

42
Q

for polarized cells, ____ are needed for transport to apical/basolateral membranes

A

different signals

43
Q

TGN structure is key to ____

A

its sorting ability

44
Q

TGN is organized into ____

A

subdomains of different proteins & lipids

45
Q

TGN enriched in _____ which serves as _____

A

-PI4P
-binding platrom for proeins involved in cargo sorting + vesciel formation

46
Q

organization of TGN subdomains dependent on ____

A

Rab GTPases

47
Q

TGN domains that sort cargo to different places have ___

A

different residues

48
Q

sorting of proteins in TGN real time method

A

live-cell imaging

49
Q

temperature block permits _____

A

synchronization of traffic moving through TGN

50
Q

GPI protein targeted to _____

A

apical surface

51
Q

adenoviruses are good for studying ____`

A

cargo targeting from the TGN

52
Q

Fluorescence microscopy helps use determine ____

A

when cargo is sorted and transport routes and which compartments are involved

53
Q

Post-golgi carriers _____ with endosomes

A

-don’t intersect

54
Q

regulated secretion are in response to ___

A

-signal (hormone/neurotransmitter)

55
Q

subset of secretory proteins (inslunin neurotransmittes) are ___ in

A

concentrated (200-400x) and stored in specialized from mature granules

56
Q

mature granules are from ___

A

maturation of immature secretory (ISGs)

57
Q

contents of MSG secreted in response to _____

A

extracellular simulates (ex. neurotransmitter)

58
Q

MSG has ____ (desc.)

A

dense core that contains high conc of protein

59
Q

proteins aggerate in response to ___ and ___ invitro

A

low pH and Ca++

60
Q

pH of TGN is ____ than golgi stack and has more ____

A

-lower
-Ca++

61
Q

when regulated secretory protein is expressed in cell that doesn’t normally express it, ______

A

protein enters regulated secretory protein suggesting that protein contain inherent targeting signals

62
Q

selection of regulatory secretory pathway: sorting by entry

A

-proteins destined for SG aggregate in special regions of TGN (high Ca++ and low PH (6.5)
-may/may not be signal mediated
-confirmation-dependent signal binds to receptor to TGN –> individual proteins/aggregates into ISGS

63
Q

selection of regulatory secretory pathway: sorting by retention

A

-non-regulatory secretory proteins removed by CCVs (clathrin coated vesicles)
-removal results in maturation into MSGs

64
Q

clathrin coats on ISG & TGN: selection method

A

selection by entry and selection by retention

65
Q

clathrin coats on ISG OR TGN: selection method

A

only one selection method is right
TGN: entry
ISG: retention

66
Q

antibodies that recognize either forms of proinsulin/insulin can be used ___

A

to determine where processing of regulated secretory protein occurs in membrane compartments

67
Q

Immuno-electron microscopy (def.)

A

gold particles coated in protein A used to detect antibody bound protein by transmission electron microcopy

68
Q

Cpn60 localizes in _____

A

secretory granules in pancreatic cells

69
Q

TM shows that proinsulin processing ____

A

happens after it leaves TGN and granules mature

z:Cell 300 (17 decks)

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