Chapter 8: endomembrane system Flashcards
biosynthetic pathways:
pathway where proteins are synthesized in the ER, modified in the golgi, and transported to other parts of the cells
secetory pathway
when proteins are discharged from the cell (from their secretory granules) into the extracellular comparment
constitutive secretion
material are discharged in a continual manner
when materials are stored in vesicles and secretory grandules and only discharged in response to a certain stimulus
regulated secretion
endocytic pathways
routes where external materials can enter the cell. frlow from the cell surface to the interior by way of endosomes and lysosomes.
how does cargo follow different pathways to different locations?
via amino acid coding signals or oligosaccharide coding signals.
An experiment that is an example of how autoradiography and electron microscopy can be used to study endosomes
a pulse chase experiment. Pulse amino acids with radiation and allow them to be taken up by the cell. After a few minutes/hours, see where they are in located in the cell “chase” to see which organelle theyre in.
the longer the chase, the further the proteins will be from the starting point (ER)
Autoradiography
a method to visualize biochemical processes using radioactive labelled Amino acids exposed to a silver photographic film.
Purpose of using GFP.
tagging GFP with a protein of interest allows the protein to emit fluoresence when created.
when the DNA for GFP and a different protein get coupled, this is called the
GFP-DNA chimera
How do you find and mark organelles to see where the GFP protein is truly located in the cell?
use mannosidase to tag the golgi, the dye is specific to golgi. if GFP and red dye overlap, you know that the protein is in the golgi.
Methods of homogenization
1) homogenizer
2) chemical lyses
3) mortar and pestle
4) glass beads
Using differential centrifugation and subcellular fractionation to study a specific organelle is an example of a
cell free system
microsomes
membranous vesicles derived from the endomembrane system (primarily the ER and golgi), some lysosomes and peroxisomes.
how would you separate the smooth and rough ER?
1) homogenization of ER
2) smooth and rough ER particle solution is placed in tubes with increasing sucrose solution
3) centrifugation
in a tube with LOW levels of sucrose: smooth Er will be in supernatant, and rough ER will be in pellet
in a tube with HIGHlevels of sucrose: smooth ER will be in the pellet and the RER will be in the supernatant.
isopyric point
region in the sucrose gradient tube where the density of the fraction = the density of sucrose (the ER microsome)
the determination of different genes that are involved in vesicular formation and the endomembrane system can be studied by:
using mutant phenotypes in an experiment
SEC12 gene:
allows for vesicles to bud off from the endoplasmic reticulum.
What would happen if you had a mutated SEC12 gene?
proteins would stay in the ER because they could not bud off, ER would swell
SEC17 gene
allows for fusion of vesicle to the golgi complex
what would happen in if you had a mutated SEC17 gene?
Vesicles would pinch off from the ER and would stay in the cytosol because they would not fuse with the golgi
How does the use of RNA interference through siRNAs affect protein translation?
siRNAs will INHIBIT translation of mRNA to protein by blocking and binding with mRNA, making mRNA appear to the ribosome as double stranded. tRNA cannot interacti with mRNA if it is hybridized with siRNA, and thus no proteins can be synthesized.
using siRNAs to stop translation produces a temporary effect called _______
protein knockdown
the network of flattened sacs of the ER is called
cisternae