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20 Cards in this Set
- Front
- Back
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What are the things to take into account when deciding what organism to get your protein from?
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amount/stability of protein
cost convenience |
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What are some things to consider when seeking to genetically manipulate protein expression?
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eukaryotes have introns
they might need post translational processing |
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describe specific activity
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the activity of the enzyme per mg of protein
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describe enzyme activity
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amt pdct formed per unit time
a unit is a mumole/min |
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What does lysozyme do in the AP exp.?
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It hydrolyzes part of the cell wall, releasing AP from the periplasmic space.
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What is DNAse needed for in the AP exp?
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DNAse breaks up DNA from broken spheroblasts, reducing the viscosity of the soln. for centrifugation.
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What is EDTA needed for in the AP exp?
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chelate Ca2+ in cell wall to facilitate lysozyme
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What is MgS needed for in the AP exp?
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Chelate excess EDTA
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What is the Michaelis-Menton hypothesis?
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ES is in rapid equilibrium with E+S
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What is the Briggs-Haldane hypothesis?
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Rate of formation of ES= rate of breakdown, where k1 and k-1>>k2
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What are the Michaelis-Menton assumptions?
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[S]>>[E] => [S] is constant
at initial velocity, [P]=0 and [S]=[S]init. rate of ES formation and breakdown are equal |
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What is the definition of Km?
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(k-1+k2)/k1
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What is the difinition of the velocity of a michaelis-menton rxn?
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v=k2[ES]
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What is the definition of Vmax?
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Vmax=k2[Et]
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What is kcat. How do you calculate it?
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Kcat is the turnover, how many moles S get converted to P by E in one minute
kcat=k2 or =Vmax/moles E |
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What is the slope of a lineweaver-burk plot? The y-intercept? The x-intercept?
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Slope=Km/Vmax
y-intercept= 1/Vmax x-intercept= -1/km |
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Define competitive, uncompetitive and noncompetitive inhibition.
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comp: I binds to E instead of S
uncomp: I binds to ES slowing production of P noncompetitive: I binds to E and ES |
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How can you discern competitive inhib. plots?
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Km changes but Vmax doesn't
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How can you discern uncompetitive inhib. plots?
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Km and Vmax change
the lines in lineweaver-burk are parallel |
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How can you discern noncompetitive inhib. plots?
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Km and Vmax change
the lines in lineweaver-burk intersect at one point, but not on any axes. |
