Endoplasmic Reticulum and Golgi Apparatus Flashcards

1
Q

What is the endoplasmic reticulum?

A

The Endoplasmic Reticulum is a continuous network of membrane tubules that is continuous with the outer layer of the nuclear membrane.

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2
Q

What is the perinuclear space?

A

The space between the double bilayer membrane of the nucleus.
It runs continuously with the endoplasmic reticulum.

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3
Q

What are the proteins made in the cytosol used for?

A

It is used to make proteins that will be used within the cell (in the nucleus, mitochondria, peroxisomes, cytosol etc.)

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4
Q

What are proteins made in the ER used for?

A

The protein create will either be used outside the cell, become embedded within the plasma membrane, or go to any of the organelles use within the secretory pathway (i.e. golgi apparatus, ER, lysosomes).

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5
Q

What are the steps involved in the secretory pathway?

A
  • mRNA will get translated via a ribosome that is found on the endoplasmic reticulum (specifically the rough endoplasmic reticulum).
  • The protein is formed and it will bud off into a vesicle to then go and join the golgi apparatus where it undergoes a maturation process (i.e sugar chains are added; processed and modified).
  • Then it buds off into another vesicle for it to be transported outside the cell or embedded to the cell’s membrane via exocytosis.
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6
Q

What are the two types of protein that can be formed on the ER?

A
  • Soluble proteins: proteins that are soluable inside of the lumen of the ER, eg. Insulin.
  • Proteins containing transmembrane domains: proteins that have a transmembrane helices that are embedded in the plasma membrane- they use their transmembrane domains to act as signals to enter the ER.
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7
Q

Name the different functions that the ER has

A
  • Protein synthesis.
  • Glycosylation: attaching sugar chains to proteins.
  • Folding and assembling newly synthesised proteins into their correct 3D structure.
  • Sit of lipid synthesis (SER) (cholesterol, phospholipids, steroids).
  • Ca 2+ imounding: storing calcium ions with the lumen of the ER and maintaining it at low levels in the cytoplasm, increasing it when required for cell signalling.
  • Also contains a large number of cytochrome p450 enzymes which are used in detoxification (if toxic molecules are eaten).
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8
Q

What cell is used to view the secretory pathway using electron microscopy and why?

A

Pancreatic Acinar cells are used as they contain lots of digestive enzymes which are stored by secretory granules and released after fusing with the enzyme when needed.

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9
Q

What is the first step of protein synthesis in regards to the ER?

A

A ribosome will attach to the mRNA strand and will start scanning untill it find the AUG codon- only after it has found the start codon it will start initiating translation.

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10
Q

What is the second step of protein synthesis in regards to the ER?

A

Proteins that are meant to be synthesised in the ER will have a hydrophobic chain of amino acids at the N-terminal, which is also known as a signal peptide as it will be used to insert the the protein within the ER.

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11
Q

What is the third step of protein synthesis in regards to the ER?

A

The signal peptide will emerge from the ribosome after translation has started, but translation is stalled untill the signal recognition particle (SRP) recognises and binds to the signal peptide.

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12
Q

What is the fourth step of protein synthesis in regards to the ER?

A

On the surface of the endoplasmic reticulum there will be a ribosome receptor, peptide translocation complex and a SRP receptor. The ribosome will bind to the ribosome receptor, the SRP will bind to the SRP receptor and the signal peptide will bind to the peptide translocation complex so the protein that is being translated can be feed through inside the lumen of the ER. Then after this is done, translation can continue.

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13
Q

What is the fifth step of protein synthesis in regards to the ER?

A

The signal peptide is no longer needed (used only to direct the mRNA to the ER) so it will be cleaved off using the enzyme signal peptidase as well as GTP being hydrolised into GDP and Pi.

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14
Q

What is the sixth step of protein synthesis in regards to the ER?

A

Translation will continue and the rest of the protein will be created within the lumen of the ER, for it to be then folded and processed into its final, functioning 3D structure using helper proteins- the protein may also then bind with other proteins to form multi-protein complexes.

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15
Q

Where does insulin synthesis occur? (soluble proteins)

A

Within beta-cells that can be found within the pancreas.

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16
Q

What is the first step of insulin synthesis?

A

A ribosome present in the cytoplasm will translate the mRNA molecule for insulin into preproinsulin, which will be then transported to the ER via a vesicle. It also has a signal sequence attached to it.

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17
Q

What is the second step of insulin synthesis?

A

In the ER, the preproinsulin will get modified and have its signal sequence cleaved off at the N-terminal, turning it into proinsulin. In addition it will also have disulfide bonds formed within it as part of it being folded into its final 3D structure.

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18
Q

What is the third step of insulin synthesis?

A

The proinsulin will then bud off into a vesicle where it will travel to the golgi apparatus to be cleaved into three segments: a chain, b chain and the c peptide- where the a chain and b chain still connected by the disulfide bonds, becoming mature insulin.
NOTE: the c peptide is used in the cell membrane for g-protein signalling.

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19
Q

What is the fourth step of insulin synthesis?

A

The mature insulin is now folded into its proper and functional 3D structure and buds off into a vesicle where it is take and fused to the cell membrane, releasing the insulin into the bloodstream.

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20
Q

What is the name of the enzyme used to cleave the signal peptide during insulin synthesis?

A

Signal peptidase.

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21
Q

What are the lengths of the different proteins in insulin synthesis?

A

 Preproinsulin – 110 amino acids long
 Signal sequence – 24 amino acid long
 Mature insulin – A chain: 21 amino acids long, B chain 30 amino acids long.

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22
Q

How many modifications can a protein undergo when processed via the endoplasmic reticulum?

A

It can undergo five principle modifications

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23
Q

What is proteolysis?

A

The cleaving off (removal) of the signal peptide by signal peptidase.

24
Q

What is disulphide bond formation?

A

The addition of disulphide bonds which help stabilise tertiary and quaternary structures and can only form in the lumen of the RER- this means that disulphide bonds can only be found within secretory and membrane proteins.

25
Q

What is glycosylation?

A

Attachment of very large complex sugar structures (many monosaccharides) to proteins.

26
Q

What is de-glycosylation?

A

Some sugars are immediately cleaved after glycosylation whilst the protein is on the ER.

27
Q

Why is the protein folded and assembled in their tertiary or quaternary structures?

A

-To enable interactions with ligands – if it will be a receptor on cell surface or interact with substrates if it’s an enzyme.
-Protein disulfide isomerase (PDI) is one such foldingcatalyst; thechaperoneHsc70 is
another

28
Q

Where do most of these modifications take place?

A

Mainly in the golgi apparatus but some addition/cleave of carbohydrates and proteolysis can occur on the ER.

29
Q

What are the names of the structures that transport secretory proteins out of the cell?

A

Porosomes.

30
Q

How is the vesicle formed when its transporting the protein from the ER to the Golgi apparatus?

A

Part of the ER membrane is cleaved off forming a transport vesicle.

31
Q

What is the name of the processing of proteins into vesicles?

A

Sorting and packaging.

32
Q

What is the names of the coating proteins that surrounds the vesicle?

A
  • COPI: vesicles that go to the ER.

- COPII: vesicles that go to the golgi apparatus.

33
Q

What do the proteins that use the secretory pathway have to follow?

A

Signal Sequence.

34
Q

What is another method of molecule transport from the ER?

A

It uses membrane contact sites- this is where the membrane of the ER and other close organelles are held close enough for lipids and small molecules to be transported through them.

35
Q

What happens to the proteins that do not pass the cell’s own mechanism of quality control?

A

Proteins that fail any one quality check will not be exported from the ER and are transported
back to cytoplasm and degraded there instead.

36
Q

What are the different checks that occurs during quality control?

A

-Proteolysis taken place correctly: however, some proteolysis take place in Golgi.
-Protein underwent Glycosylation and initial steps of De-glycosylation must have taken
place.
-Disulphide bond formation: protein has to be in its proper active configuration.
-Folding and assembly into complex (multi-protein).

37
Q

What is the first step involved in creating the COPII protein coat for ER transport?

A

First, sar1 (an enzyme) hydrolyses GTP into GDP and Pi to generate energy for the process.

38
Q

What is the second step involved in creating the COPII protein coat for ER transport?

A

Sec 23/24 protein complex then binds and interacts cargo that is either specific receptors or have transmembrane domains and bind to the soluble proteins.

39
Q

What is the third step involved in creating the COPII protein coat for ER transport?

A

Sec 13/31 comes from the cytoplasm and clusters on the membrane. They polymerise causing deformation of the membrane. Eventually, as they continue to polymerise the deformity causes the vesicle to pinch off.

40
Q

What is the structure of the golgi apparatus?

A

Consists of flattened membrane-enclosed sacs (cisternae) and is located near the cell nucleus (in animal cells, can be found near the centrosomes).

41
Q

What are the names of the different parts of the golgi apparatus?

A
  • cis face: this is the entry point of molecules and is the part closest to the ER. Cis-golgi network.
  • trans face: this is the exit point where proteins are exported to various destinations, pointing towards the plasma membrane. Trans-golgi network.
  • medial stacks: the middle stacks of golgi. Contains cis- cisternae, medial-cisternae and trans-cisternae.
42
Q

What are the roles of the golgi apparatus?

A

-Protein modification:
*de-glycosylation catalysed by glycosidase.
*attachment of new sugars catalysed by
glycotransferase.
*removal of sulfate groups by sulfatases.
*protein cleavage by proteases.
-Lipid synthesis:
*sphingolipids completed in Golgi e.g. Sphingomyelin,
glucosylceramide (types of sphingolipids).
-Sorting proteins and lipid to different places in the cell:
*secretory granules
*plasma membrane
*basolateral versus apical membrane
*endosomes
*lysosomes

43
Q

What are the two types of exocytosis?

A

Constitutive and regulated exocytosis pathways.

44
Q

What is constitutive exocytosis pathway?

A

When there is a steady stream of vesicles from the golgi to the plasma membrane.
Supplies newly made lipids and proteins to the plasma membrane which is used for plasma membrane growth (this is useful for when the cells enlarge during G1).
Carries proteins to the surface of the cell to be released outside- some becoming peripheral proteins and some are also incorporated into the extracellular matrix.

45
Q

What is regulated exocytosis pathway?

A

This pathway is for cells that specialise in secretion.
Specialized secretory cells produce large quantities of particular products, such as hormones, mucus, or digestive enzymes, which are stored in secretory granules for later release.
These vesicles bud from the trans Golgi network & accumulate near the plasma membrane
*An extracellular signal will stimulate them to fuse with the plasma membrane & release their contents to the cell exterior.

46
Q

What happens to the vesicle membrane when it fuses with the plasma membrane?

A

It becomes apart of the plasma membrane.

47
Q

What happens to the surface area of the membrane after the vesicle membrane is fused with it?

A

It’s surface area momentarily increase but due to the other areas of the membrane being removed by endocytosis, the rate of adding to the membrane and removing from the membrane is almost equal (exocytosis rate=endocytosis rate).

48
Q

What is the SREBP cycle used for?

A

Sterol regulatory element-binding proteins are transcription factors and the cycle is used to regulate cholesterol.

49
Q

What are the names of the 3 proteins embedded in the ER membrane that are important the SREBP cycle?

A
  • Insig
  • SCAP
  • SREBP
50
Q

How is SREBP synthesised as?

A

As an inactive precursor protein with 2 transmembrane domains that anchor it to the membrane of the ER.

51
Q

What happens to the 3 proteins on the ER when the cholesterol levels are high?

A

The remain in the ER.

52
Q

What happens to the 3 proteins on the ER when the cholesterol levels are low?

A

1) The Insig-SCAP-SREBP complex separate
- The SCAP-SREBP complex move through a particular
pathway (COPII mediated vesicular transport) to the Golgi apparatus
- At the same time Insig is degraded
2) 2 proteases cleave the SREBP transcriptional factor
- Both act sequentially to relieve a part of the SREBP from the Golgi membrane
- The relived part (n-SERBP) migrate to the nucleus
3) In the nucleus it will bind to specific target genes & activate their transcription
4) Those target genes are for:
- Cholesterol synthesis
- LDL receptors

53
Q

How is the system of maintaining levels of cholesterols regulated?

A

Proteases only present in the Golgi apparatus
Regulated release of insig and regulated transport of the SREBP from the ER to the Golgi ensures cholesterol only happens when cholesterol levels are low.
When cholesterol levels go back to normal, the system will stop and prevent accumulation of product.

54
Q

What products can Insig also activate the transcription for?

A

Cholesterol and Insulin.

55
Q

What are the main methods used to study golgi transport?

A
  • Electron microscopy
  • Light microscopy
  • X-ray crystallography