L11

Question 1

what does an uncompetitive inhibitor do to an enzyme

Answer 1

binds the ES and stops transition to EP

Question 2

what part of the enzyme can you speed up

Answer 2

the amount of time it takes to find the substrate, but not the other time it takes to make the products

Question 3

competitive Vmax and Km

Answer 3

Vmax same
Km higher

Question 4

non-competitive inhibitor Vmax and Km

Answer 4

Vmax lower
Km same

Question 5

uncompetitive inhibitor Vmax and Km

Answer 5

Vmax lower
Km lower

Question 6

what is the concentration of an enzyme called

Answer 6

activity

Question 7

what are the units of activity

Answer 7

U/ml or U/ul

Question 8

when you multiply activity by volume of enzyme, what is that

Answer 8

total activity

Question 9

whats the potency of an enzyme or weight of protein called

Answer 9

specific activity

Question 10

what are the units of specific activity

Answer 10

U/mg

activity/ protein concentration

Question 11

how to calculate specific activity

Answer 11

units of enzyme / amount of protein

Question 12

how to calculate enzyme activity

Answer 12

units of enzyme / volume

Question 13

how to calculate total activity

Answer 13

activity x volume

Question 14

when specific activity goes up, what does that indicate?

Answer 14

that we have purified our enzyme away from other proteins

Question 15

when activity goes up, what does that mean

Answer 15

just that the concentration went up, doesn’t really tell us if we’ve purified

Question 16

what enzymes are used for protein extraction

Answer 16

cytoplasmic enzymes

Question 17

what enzymes break down proteins

Answer 17

proteolysis

Question 18

what should be done to make sure no enzymes or proteins die once extracted from the cell (3)

Answer 18

ph changes
heat denaturation
oxidation

Question 19

assays should be: (4)

Answer 19

sensitive
specific
rapid
quantitative

Question 20

what kind of purification is electrophoresis

Answer 20

charge

Question 21

what kind of purification is gel electrophoresis

Answer 21

size

Question 22

what must the protein be in order for it to be centrifuged

Answer 22

soluble

Question 23

what is the supernatant

Answer 23

the protein at the top of the centrifuge tube

Question 24

whats a common and reversible purification method

Answer 24

salting out

Question 25

what kind of purification is salting out

Answer 25

solubility

Question 26

what is dialysis

Answer 26

de salting

Question 27

what is dialysis good for? bad for?

Answer 27

great for separating proteins from salts bad for separating proteins based on size

Question 28

what is chromatography

Answer 28

techniques to separate mixtures based on physical properties such as size or charge

Question 29

whats the stationary phase

Answer 29

a substance that the compounds to be separated pass by or interact with

Question 30

whats the mobile phase

Answer 30

the carrier for the compounds to be separated

Question 31

5 steps to column chromatography

Answer 31

pouring packing loading running/eluting collecting

Question 32

what makes up stationary phase in size exclusion chromatography

Answer 32

porous beads

Question 33

whats the volume of the column

Answer 33

Vt

Question 34

volume outside the beads

Answer 34

Vo

Question 35

volume inside the beads

Answer 35

Vi

Question 36

volume at which sample is eluted

Answer 36

Ve

Question 37

partition coefficient

Answer 37

fraction of the pores within the beads available to the sample

Question 38

if Kav is 1, what does that mean

Answer 38

beads are fully accessible

Question 39

if Kav is 0, what does that mean

Answer 39

always excluded by beads

Question 40

what will be the fastest traveller

Answer 40

Vo