Protein Purification - Zaidi

Question 1

What is protein purification and why is it used?

Answer 1

isolation of a single protein form thousands of others in a cell; used to study the structure and function of individual proteins

Question 2

How does protein purification work?

Answer 2

uses recombinant DNA technology to over express a protein and then it is purified as an endogenous protein

Question 3

In regards to protein purification, what is something you usually need to do before purification?

Answer 3

sub-cellular fractionation which reduces the complexity of the material

Question 4

What is the tissue of sub-cellular fractionation?

Answer 4

mechanical bleeding

Question 5

In regards to sub-cellular fractionation, what is homogenate?

Answer 5

suspension of different cell types

Question 6

In regards to sub-cellular fractionation, what does centrifugation do?

Answer 6

separateds different cell types based on size and density

Question 7

In regards to sub-cellular fractionation, what does ultracentrifugation do?

Answer 7

separates organelles

Question 8

In regards to cell fractionation by centrifugation, what do the pellets form varying speeds contain?

Answer 8

low-speed: whole cells, nuclei, cytoskeletons

medium-speed: mitochondria, lysosomes, peroxisomes

high-speed: microsomes, small vesicles

very-high speed: ribosomes, viruses, large macromolecules

Question 9

What makes up the synaptosome?

Answer 9

The presnyaptic and postsynaptic areas and the synaptic cleft in between them

Question 10

In regards to different matrices for column chromatography, what are the differences between the requirements for ion-exchange, gel-filtration, and affinity chromatography?

Answer 10

ion-exhange: charge

gel-filtration: molecule size

affinity: ligand binding

Question 11

PMCA purification from SPMs is an example of what type of chromatography?

Answer 11

affinity