Topic 4 Purification Flashcards
1
Q
Formula
Fraction of binding sites
A
θ = [PL] / ([PL]+[P])
or
θ = [L] / ([L] + Kd)
2
Q
Kd formula
A
Kd = [P][L] / [PL]
3
Q
Structure of Edman reagent
A
Ph-N=C=S
4
Q
Name of Edman degradation reagent
A
phenylisothiocyanate (PITC)
5
Q
Where does the Edman reagent react ?
A
The N terminus
6
Q
Size limit to Edman degradation
A
approx 100 amino acids
7
Q
AAs destroyed when protein is boiled in acid
A
Destroys Trp
Hydrolyses:
Asn -> Asp
Gln -> Glu
8
Q
Trypsin cutsite
A
After K, R
but not after Pro
9
Q
Chymotrypsin cutsite
A
after FYW
but not after Pro
10
Q
Elastase cutsite
A
after AGSV…
but not after Pro
11
Q
V8 protease cutsite
A
after DE
12
Q
Asp-N protease cutsite
A
before DE
13
Q
CNBr cutsite
A
After M
14
Q
Protease reagents (6)
A
Trypsin, Chymotrypsin, Elastase V8 Protease Asp-n protease CNBr
15
Q
What is a paralog ?
A
It is when an enzyme replicates and mutates within the dna of one species
