Lecture 11&12 - Membrane bound organelles and Protein sorting Flashcards

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1
Q

internal organelle evolution book section

A

One suggested pathway for the evolution of the eukaryotic cell and its internal membranes As discussed in Chapter 1, there is evidence that the nuclear genome of a eukaryotic cell evolved from an ancient archeaon. For example, clear homologs of actin, tubulin, histones, and the nuclear DNA replication system are found in archaea, but not in bacteria. Thus, it is now thought that the first eukaryotic cells arose when an ancient anaerobic archaeon joined forces with an aerobic bacterium roughly 1.6 billion years ago. As indicated, the nuclear envelope may have originated from an invagination of the plasma membrane of this ancient archaeon—an invagination that protected its chromosome while still allowing access of the DNA to the cytosol (as required for DNA to direct protein synthesis). This envelope may have later pinched off completely from the plasma membrane, so as to produce a separate nuclear compartment surrounded by a double membrane. Because this double membrane is penetrated by nuclear pore complexes, the nuclear compartment is topologically equivalent to the cytosol. In contrast, the lumen of the ER is continuous with the space between the inner and outer nuclear membranes, and it is topologically equivalent to the extracellular space
(slide 4)

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2
Q

What is the function of peroxisomes

A

Fatty acid beta oxidation

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3
Q

WHAT IS IN THE ENDOMEMBRANE SYSTEM

A

Endoplasmic reticulum, lysosomes, Golgi, Secretory vesicle

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4
Q

What is the benefit of an endomembrane system

A

molecules can get from one to the other without having to cross a membrane

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5
Q

How are organelles in the endomembrane system connected

A

protein trafficking system

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6
Q

what is the entry point for newly synthesised proteins in the endomembrane system

A

Endoplasmic reticulum

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7
Q

What is a translicon

A

Protein conducting channel

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8
Q

How long are ribosomes on the RER

A

Until protein has been synthesised, then dissociate into big and small subunits and search for mRNA

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9
Q

Where does N-linked glycosylation occur

A

ER

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10
Q

What is N-linked Glycosylation + mechanism

A

the attachment of an oligosaccharide, a carbohydrate consisting of several sugar molecules, sometimes also referred to as glycan, to a nitrogen atom

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11
Q

What is the most important organelle in protein synthesis

A

ER - mis-folded or mis-assembled dont leave ER - folding and quality control - disulphide bond formation

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12
Q

Calnexin binding

A

Calnexin binds to glycans found in membrane proximal domains while the soluble calreticulin associates with glycans within the ER lumen

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13
Q

What is a reducing environment

A

An area where few disulphide bonds form

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14
Q

Describe the Golgi network

A

Slide 11

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15
Q

O linked glycosylation

A

O-linked glycosylation is the attachment of a sugar molecule to the oxygen atom of serine (Ser) or threonine (Thr) residues in a protein. O-glycosylation is a post-translational modification that occurs after the protein has been synthesised

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16
Q

Why are lysosomes degradative organelles

A

contain enzymes called acid hydrolases, work at lower pHs

17
Q

How many different protein molecules are in a eukaryotic cell

A

10 billion

18
Q

Where does protein sorting start?

A

Cytosol

19
Q

Where does the sorting signalling tend to be to import into mitochondria

A

N-terminus of proteins (amphipathic)

20
Q

What key do proteins that are imported into the nucleus (signalling) have

A

Patches of positively charged amino acids

21
Q

What is nuclear targeting

A

Proteins are directed into the nucleus using patches of positively charged amino acids

22
Q

Why is the nuclear membrane described as contiguous

A

Envelope - two membranes, and continuous with Endoplasmic reticulum

23
Q

What is the function of nuclear pores

A

All movement of molecules into and out of the nucleus occurs through nuclear pores
(slide 26 - nuclear pore complex)

24
Q

How does protein import through nuclear pore complex occur

A

Nuclear import receptors recognise NLS’ on prospective nuclear protein
Complex of receptor and protein with NLS guided to nuclear pore by fibrils
Binding of nuclear protein to pore opens pore
Active transport into nucleus (together with receptor

25
Q

What do GTPases do

A

Work as molecular switches - work in GTP bound state or GDP bound state in different conformations

26
Q

How does GTP convert to GDP and reverse

A

Hydrolysis eg. Ran GAP (GTPase activating protein) activates Ran-GTP to hydrolyse to Ran-GDP

(reverse Ran-GEF - Guanine nucleotide Exchange Factor) facilitates the exchange of GDP (removed completely) for GTP

27
Q

Where are Ran-GEFs located

A

Nucleus

28
Q

Where are Ran-GAPs located

A

Cytoplasm

29
Q

Watch animation 15.1

A

Nuclear import/export (slide 13)

30
Q

What are some key features of mitochondrial targeting sequences

A

Always at the N-terminus
Varies in size from 20-80 amino acids
Multiple positively charged amino acids make an amphipathic a-helix
(Watch Movie 15.2)

31
Q

How do proteins get into mitochondria (channels)

A

Enter TOM and TIM (translocation channels - extremely specific) (Outer and inner membranes)

32
Q

How do proteins enter channels

A

proteins need to be unfolded proteins and associate to chaperone proteins to keep them unfolded to be imported (active process as well as gradient creation)

33
Q

Why are mitochondria negatively charged

A

Pumping out protons in generation of ATP

34
Q

Where are mitochondrial proteins located

A

Outer membrane, Inner membrane, Inter-membrane space or Matrix (own genome, and protein synthesis machinery)

35
Q

How does a protein target to the ER

A

Hydrophobic signal sequence (mostly at N terminus)

36
Q

Describe ER targeting and translocation

A

Ribosome binds to RNA causing translation

Binds to SRP (Signal Recognition Particle) to signal peptide causes a pause in translation

SRP-bound ribosome attaches to SRP receptor in ER membrane,

Translation continues and translocation begins

SRP and SRP receptor displaced and recycled

Pushing of protein into lumen of ER, negatively charged n-terminus pushed through translicon - cotranslational translocation

Protein refolds once it has entered endoplasmic reticulum
(movie 15.4 and YT link slide 21)

37
Q

Ribosome-nascent chain complex

A