Endoplasmic reticulum Flashcards
What is the primary role of the secretory pathway?
Primarily the transport of modified proteins from the inside of the cell to the outside.
What are the three mechanisms of protein transport?
Nuclear pores
Vesicles
Transmembrane
What is the ER made of?
Reticulum: network of membrane tubes
Plus outer nuclear envelope
Closed membrane: contents separate from cytoplasm.
What are the key aspects of ER structure?
Cisternae are long flattened, sac like, unbranched tubules.
Fluid filled lumen or the cisternal lumen where biochemical reactions occur.
It is a closed organelle that provide a unique environment.
What are the main differences between the rough and smooth ER
RER: important for secretary pathway and protein biosynthesis - folding proteins, regulating misfolded proteins and responding to cellular stress. Tissue types with high protein demand will have cells with lots of RER.
SER: main role is the biosynthesis of phospholipids and cholesterol and the repair of membranes.
Hepatocytes for example have a lot of SER
What is translocation?
Movement of proteins across a membrane.
What are the different components of the secretory pathway / endomembrane system?
- ER
- Golgi
Then either transport vesicles through cytosol to plasma membrane
Or to endosomes and lysosomes
What organelle is required in larger quantities in cells that require the manufacturing and secretion of a lot of proteins, why?
Mitochondria, to provide the necessary energy.
What is co-translocation?
When proteins are translated and translocated at the same time at the RER
Which ribosomal subunit is attached to the RER, whats the other one doing?
The large subunit is attached and the small subunit exists separately and are recruited as they needed for translation.
How is the correct ribosome get targeted to the RER for a specific process.
A signal sequence of amino acids on the RER act as a ‘postcode’ for transport destinations. The ribosome is connected to an mRNA transcript that recognises the signal sequence.
What is a translocon?
A channel protein complex that transverses ER the membrane and exports and imports proteins to the ER. It has a roughly hour glass shape with a pore in the middle for storing proteins. The is a ‘plug’ region of the complex at the ER interface to prevent the leakage of ions during translocation of a protein substrate.
What do the signal recognition particle and receptor proteins do?
They act together during co translocation
The SRP interacts with the newly synthesised polypeptide/ribosome to transport the protein being translated to the translocon. The SRP briefly pauses translation while it finds and binds to the SRP receptor in the ER membrane. The receptor recruits the translocation channel and translation continues.
How does a protein pass through a channel/translocon during co translational translocation?
The signal sequence of a soluble protein attaches to the channel and then the protein passes through with the aid of a chaperone protein (eukaryotes: BIP, prokaryotes: Kar2) then an enzyme (signal peptidase) cleaves the signal sequence off the protein releasing the protein into the ER lumen. Thee protein can now be folded for its function.
How does a protein pass through a channel/translocon during post translational translocation?
This is done independently of SRPs and receptors. Chaperone proteins still pull the protein through but due to the lack of the aid of SRP and receptor proteins, there are other transmembrane proteins that help the process occur.
In what conditions can disulfide bonds form between which amino acids? Where is this found commonly?
In an oxidative environment such as the ER lumen between cysteines.
What is the protein disulfide isomerase?
An ER lumenal oxidoreductase (enzyme) that catalyses redox reactions that alter the position of disulfide bonds on proteins.
What is glycosylation?
A method of modifying proteins in the ER lumen by adding oligosaccharides (carbohydrates) as the protein passes from the cytosol to the ER lumen. Only extracellular parts of the protein are glycosylated. Glycosylated proteins function as cell receptors.
How does glycosylation occur? What type of proteins does it make?
Transmembrane dolichol proteins that are attached to a lipid linked oligosaccharide protruding into the ER lumen will transfer the oligosaccharide to the growing peptide chain attached to the translocon. The enzyme oligosaccharide protein transferase facilitates the transfer.
Glycoproteins that mostly act as cell receptors.
How is protein export from the ER regulated?
Misfolded proteins are held in the ER by chaperone proteins until they are either correctly folded or degraded.
How in vague terms do protein quality control checks occur
Chaperone cycle checks -> trimming proteins trim the oligosaccharide -> checks for functionality -> if not there is more trimming or degradation.
What is ERAD and how does it work?
ER associated degradation, misfolded proteins either from ER lumen or cytosol will be recognised. From the ER the misfolded protein will be exported via retro translocation by the translocon. Ubiquitin ligase complexes that are associated with the ER lumen will attach ubiquitin to the misfolded protein which targets the protein for degradation by a proteosome .
What is the UPR and what does it do in the ER?
The unfolded protein response is mechanism which deals with a backlog of unfolded proteins either by signaling apoptosis or fixes the backlog. The build up of unfolded proteins will trigger the upregulation of chaperone proteins to fold the proteins and it will trigger expansion of the ER to create a dilution effect to deal with the excess proteins. Another part of the response is the triggering of transmembrane receptors which signal there is a ‘problem’. This is a feedback response.
What is Ire1 and what does it do?
A transmembrane protein that is part of the UPR. In its inactive state it is monomeric and unphosphorylated, it also has BIP protein bound to it. When its activated by unfolded proteins, BIP dissociates allowing it to dimerises and undergo trans autophosphorylation (where 1 protein phosphorylates the other). When active it splices an intron of a specific mRNA transcript turning into a protein that is a transcriptional activator of chaperone protein genes and it also destroys certain mRNAs ultimately reducing the load of misfolded proteins.
