S1: Fate of Newly Synthesised Proteins Flashcards Preview

MCBHD- Molecular and Cellular Basis of Health and Disease > S1: Fate of Newly Synthesised Proteins > Flashcards

Flashcards in S1: Fate of Newly Synthesised Proteins Deck (40)
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1
Q

What are free ribosomes?

A
  • Soluble proteins for release into the cytoplasm are synthesised by free ribosomes
  • Intitial codons in mRNA do not code for hydrophobic amino acids
2
Q

What are bound ribosomes?

A
  • Proteins for secretion from the cell or for incorporation into membrane or lysosomes are synthesised by membrane bound ribosomes
  • Initial codons in mRNA code for a short sequence of hydrophobic amino acids called a signal sequence
3
Q

What is a signal sequence?

A

The initial codons (so at the N-terminus, the first part of the protein to be synthesized) in the mRNA code for a short sequence of hydrophobic amino acids, called a “signal sequence”.

4
Q

What determines whether the ribosome that translates the polypeptide will be free or bound?

and How?

A

Signal sequence

The signal sequence allows a channel in the ER the open up and the signal remains bound to channel as rest of polypeptide enters into the ER lumen. When it has fully entered, the signal sequence is cleaved off and released.

5
Q

What are internal hydrophobic sequences?

A

They are additional sequences of codons in the mRNA that code for hydrophobic amino acids.

These sequences of AA’s get ‘stuck’ in the membrane as the protein is fed through (the channel into the lumen) so they become membrane proteins.

6
Q

What is the ‘nuclear localisation sequence’?

A

These are patterns of codons that code for basic amino acids

They will appear on proteins that are destined for the nucleus

7
Q

What are ‘mitochondrial target signals’?

A

They are alternate patterns of hydrophobic and basic amino acids. This is because while mitochondria do have their own ribosomes and make some of their own proteins, the majority of mitochondrial proteins are synthesised in the cytosol and migrate to the mitochondria.

8
Q

Organelles involved in protein synthesis

A

The mRNA leaves the nucleus and is translated on a ribosome on the rough ER (for proteins that will be secreted or part of membrane), it then will move to the Golgi apparatus where it’ll be packaged into vesicles, these will then fuse with the plasma membrane and empty the contents into the ECF, this is secretion or it will add new membrane to the surface of the cell, including some the hydrophobic proteins with it.

There will also be messages translated in the cytosol, proteins of which will be used in the cell.

9
Q

What are the two types of ER?

A

Rough ER

Smooth ER

10
Q

Describe characteristics of RER

A
  • Synthesis of proteins for subsequent packaging and secretion from the cell or for insertion into intracellular structures such as other membranes
  • It is involved in the initial steps of glycosylation (joining carbohydrates onto proteins)
  • Site of disulphide bond formation (which is important for the folding of the protein into its tertiary structure, chaperones help)
11
Q

Describe characteristics of smooth ER

A
  • Modification of newly synthesised proteins –> addition of carbohydrate, phosphate and lipid group
  • In the liver the SER is responsible for detoxification of foreign compounds such as drugs and environment pollutants (making drugs more soluble)
12
Q

Describe the formation of disulphide bonds in the ER

A
  • On the surface of the ER is where there is formation of disulphide bonds between cysteine residues
    (S-H group in side chain)
  • The formation of disulphide bonds is catalysed by the enzyme protein disulphide isomerase (PDI) which resides in the ER
  • PDI is involved in breaking and reforming the disulphide bonds until the protein is the correct structure

-

13
Q

What group is PDI (protein disulphide isomerase) part of?

A

Folding chaperones

14
Q

What do folding chaperones do?

A

The polypeptide chains interact with chaperones, which promote the folding of proteins to the correct structure. Folding chaperones also allow re-folding if it has folded into an inappropriate structure.

15
Q

List factors that can cause protein misfolding

A
  • Cells are exposed to elavated temperatures resulting in increased expression of some chaperones (heat shock proteins) so proteins are more likely to misfold
  • HSP are also expressed involved more in stressed individuals
16
Q

What happens is the degradative process for misfolded proteins becomes overwhelmed?

A

Protein aggregates can accumulate causing cellular dysfunction and cell death (apoptosis)

17
Q

Explain the destruction for unwanted proteins

A
  • Unwanted proteins are marked for degradation by tagging lysine residues (on the protein to be destroyed) with a polymer of ubiquitin (polypeptide that acts as an tag)
  • Ubiquitin marks the protein so it can be recognised by proteasomes
  • The individual amino acids are cleaved off as the protein goes through the hollow core of the proteasomes
18
Q

Why are ubiquitin ligases important?

A

Ubiquitin ligases are very important in the body, there are about 600 ligases encoded by the human genome (many involved in inflammatory, cardiovascular, metabolic diseases or cancer)

19
Q

What are proteasomes?

A

Cylindrical complexes of protein with a hollow core in which degradation occurs by an energy dependent process

They destroy proteins that have been synthesised by the cell itself, not external ones

20
Q

Describe the destinations of proteins synthesised within the ER

A
  • Proteins synthesised within the ER are transported to the golgi apparatus
  • Portions of the ER are pinched off, forming transport vesicles which carry proteins to the golgi apparatus
21
Q

What is vesicular trafficking regulated by?

A

Vesicular trafficking is vehicle transport from one site in the cell to another

It is regulated by specific coat proteins on the vesicle surfaces which are regulated by small GTPase.

22
Q

What does membrane fusion between transport vesicle and golgi apparatus depend on?

A

SNAREs

They allow the protein to migrate to the appropriate site - they site on the cytosolic face of the vesicle

23
Q

What are the two fates on the genes products?

A
  1. Secreted protein
  2. Membrane protein
  • Secreted protein exons do not contain a membrane spanning region, so they are just secreted
  • Membrane protein, exon 3 has a membrane spanning region so it gets locked in the membrane and vesicle end up as part of the membrane
24
Q

Explain how there can be different outcomes from one transcript

A

e.g. with antibodies it needs to have a membrane bound and secreted form.

Processing of the transcript therefore determines the genes fate - not all the exons would have to be used so it depends on the splicing

25
Q

What is the structure of the golgi apparatus?

A
  • It is a set of flattened bags
  • The surface closest to the nucleus is called the ‘cis’ face where the vesicles enter the golgi apparatus
  • ‘Trans’ is the part that faces the plasma membrane this is where vesicles leave the golgi apparatus
26
Q

Function of the Golgi Apparatus

A
  1. Further modifications of newly synthesised proteins particularly by addition of carbohydrate residues (glycosylation)
  2. Synthesis of glycolipids (glycosylation of lipids)
  3. Proteolytic conversion of proforms of proteins into mature forms
  4. Packaging of proteins destined for export into secretory vesicles or granules
  5. Targeting of proteins (via vesicles) to lysosomes
27
Q

Where does protein glycolysation start and end?

A

Protein Glycolysation Starts in the ER and is completed in the Golgi Apparatus

Carbohydrate addition is initiated in the ER, concomitant with protein synthesis.

28
Q

What does glycolysation depend on?

A

Many secreted and membrane proteins are glycosylated.

Glycosylation depends on recognition of specific sequence patterns (sequence of amino acids) in the target protein.

29
Q

What are the two common types of glycoproteins?

A

N-linked glycoproteins(carbohydrate built on asparagine)

O-linked glycoproteins (carbohydrate built on serine or threonine)

30
Q

Give examples of N-linked glycoproteins

A

Serum glycoproteins involved in haemostasis (blood clotting) and most membrane proteins

31
Q

Explain the structure of N-linked glycoproteins

A

Branched carbohydrate chains attached to asparagine residues when they are in the following order : Asn - X - Ser/Thre

Typical sugars involved in these structures are N-acetylglucosamine, mannose, galactose and sialic acid

Carbohydrates are important receptors for a number of ligands e.g. sialic acid is the receptor for the flu virus haemagglutin (a protein that binds to sialic acid residues).

32
Q

Explain the structure of O-linked glycoproteins

A

They typically have shorter chains (compared to N) attached to serine or threonine residues

Typical sugars are N-acetylgalactosamine, galactose, sialic acid and fucose

carb built on serine or threonine

33
Q

Examples of O- linked glycoproteins

A

Mucins and blood group antigens (ABO groups are determined by glycosyltransferase genes, the oligosaccharidechain is found on both glycoproteins and glycolipids)

Mucins provide a protective barrier on epithelial surfaces

34
Q

Explain protein maturation in the golgi apparatus

A
  • Most biologically active proteins are synthesised initially as large precursor molecules
  • Proteolytic cleavage occurs at few specific sites where specific enzymes in the Golgi cleave precursor proteins to their biologically active form
35
Q

How can you generate multiple hormones from one polypeptide?

A

POMC cleavage

36
Q

What are the two pathways for protein secretion?

A

Constitutive pathway: As the vehicle containing protein becomes mature and then continuously moves towards the plasma membrane and continuously fuse releasing their contents e.g. albumin from hepatocytes

Regulated pathway: Protein accumulates in vesicles but will not be released/fuse until controlled factor reaches a certain level. It is controlled by Ca2+ dependent signalling.

37
Q

How are lysosomal enzymes synthesised?

A
  • lysosomal enzymes may be converted from inactive precursors to active enzymes in a process acting in the golgi
  • some lysosomal proteins are autocatalytic (it can cleave other molecules and make them into active protease) whereas other enzymes require an additional protease

Activation involves proteolytic removal of part of the polypeptide chain

38
Q

What happens to proteins glycosylated with residues of mannose-6-phosphate?

A

Proteins glycosylated with residues of mannose-6-phosphate will leave the Golgi in transport vesicles that eventually fuse with lysosomes.

39
Q

List some post translational modifications

A

1) Glycosylation (addition of carbohydrate groups)
2) Phosphorylation (addition of phosphate groups) – a key way in which proteins are regulated
3) Acetylation (addition of acetyl groups)
4) Methylation (addition of methyl groups)
5) Lipoprotein formation (addition of lipid groups)

40
Q

Give an example of regulating protein activity by phosphorylation

A

PHOSPHORYLASE (releases glucose-1-phosphate from glycogen)

Phosphorylase is usually inactive in cells (as phosphorylase b). Phosphorylase kinase gets activated after exposure to glucagon and adrenaline and it converts phosphorylase b to phosphorylase a which is active. Phosphorylase a can then perform its function.

To switch off phosphorylase a, protein phosphatase (activated after exposure to insulin) removes the phosphate and converts it back to phosphorylase b (inactive).