Test 2 SG Flashcards
As percent recovery drops, what happens to fold purification?
It increacses
What does the highest fold purification step denoted by?
It’s the most effective step, calculated by specific activity
What does the lowest % recovery denoted by?
It is the most costly step, calculated by total activity.
In cation exchange, what is the charge of the Resin?
- charge, capturing + charged ions.
What is the elution order for cation exchange?
-charge, neutral, + charge
What is the charge of the resin in anion exchange?
It is a + charged resin, attracting - charged ions.
What’s the elution order of the anion exchange?
+charge, neutral, -charge.
What does SDS stand for and what does it do?
Sodium dodecyl sulfate, makes the proteins uniform in - charge and uncoils them.
What does trypsin cleave?
+ charged A.A (Lys and Arg)
What does Chymotrypsin cleave?
Aromatic A.A. (Phe, Tyr, Trp)
What does CNBr cleave?
Internal Methonines
What does Edman digestion create?
A one by one cleave of all the A.A’s, known as the PTH derivative.
What is activation energy represented by?
Delta G not plus, plus
What is free energy represented by?
Delta G not
What is the free energy equation?
-RTLnKeq
What does K represent?
The rate constant
What does Km represent?
The Michalis constant
What is the M-M equation?
V initial = Vmax*(S)/Km + (S)
What is the Lineweaver burke equation?
1/V = Km/Vmax*1/S + 1/Vmax
What is Kcat?
The Vmax relationship to turnover number of enzyme. = Vmax/Et
What does competitive inhibition do to Km and Vmax?
Km increases, no change in Vmax
What does non-competitive inhibition do to km and vmax
Lowers vmax, no change in km
What change does mixed inhibition have on Km and Vmax
Changes both Km and Vmax
What change does uncompetitive inhibition have?
Decreases both km and vmax.
What is an irreversible inhibitor also known as?
Trojan horse or suicide substrate
What does homotrophic effect do?
Several identical molecules bind to the protein
What does heteroptrophic effect do?
Several different molecules bind to the protein
What does R denote? T?
R=Active, more tightly bound. T- inactive, less tightly bound.
What does L denote? What does a higher L mean?
L denotes the ratio of T/R. The higher the L value, the higher the amount of T, the more inactive and more sigmoidal it is.
What does R denote? What does a higher R mean?
R denotes the rate of r/rate of t. The higher the R, the most active it is, and the more hyperbolic it is.
What three amino acids are phosphorylated? Why?
S (serine), T (threonine), and Y (tyrosine). They all feature an OH group.
What is a zymogen?
An inactive precursor of an enzyme, becoming active from cleavage.
What are two examples of zymogens?
Chymotrypsin and pi-Chymotrypsin.
What does chymotrypsin become pi-chymotrypsin? How does pi-chymotrypsin become alpha chymotrpisin?
Chymotrypsin must be cleaved of trypsin, while pi chymotrypsin will self digest and cleave bonds.
