Last class Gel filtration PAGE SDS vs Native.

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Presentation transcript:

Last class Gel filtration PAGE SDS vs Native

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

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

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

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 =

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

The plot thickens…

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

Practical considerations Asymptote – Reproducible? Solution?

Better method: LWB plot Asymptote – Reproducible? Solution?

Better method: LWB plot

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!

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

Studying proteins How would you study a protein?

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