1. Last class
Gel filtration
PAGE
SDS vs Native

2. The MM equation Assumptions: v0 is _________ reaction rate
_____________ velocity
Measured at ____________: Formation of ES = Breakdown of ES
[S] >>>> [E]
Measured at given ________

3. What do the terms mean? v0 v0 is _________ reaction rate
Measured at given [S] and given [E]

4. What do the terms mean? Vmax
Vmax is _____________________ reaction rate
Reaction rate when [ES] = _________
Vmax = k2 * __________
Can it ever be practically achieved?

5. What do the terms mean? Km
V 0 = V Max [S] S + K m
S + K m = V Max [S] V 0
S + K m = V Max [S] 0.5 ∗ V Max
S + K m = 2[S]
K m =

6. What do the terms mean? Km
Km is _____________ of enzyme for substrate
It is not 2 * Vmax!

7. The plot thickens…

8. Practical considerations
How to find Km and Vmax for an enzyme?

9. Practical considerations
Asymptote – Reproducible?
Solution?

10. Better method: LWB plot
Asymptote – Reproducible?
Solution?

11. Better method: LWB plot

12. Lab Fill out tables correctly!
Change [E]  Effect on rate of reaction?
Calculate Km for different substrates
- Get rate over time using different [S]
- Get v0 for each reaction rate (how?)
- MM and LWB plots
- Look over data!

13. Studying proteins
What are things that you might want to know about a protein?

14. Studying proteins
How would you study a protein?

15. Studying proteins
How to tell what a protein is need for?