Determine the km of mutant proteases

Direction depending on the needs of the animal
May 2, 2020
Solution-Daughter with normal color vision
May 2, 2020

Determine the km of mutant proteases

You have successfully developed two new inhibitors (Alice-1 & Alice-2) for the mutant peptidase mentioned in question To understand how the inhibitors work, you carried out enzyme assays measuring the ability of the mutant peptidase to cleave the substrate in the absence and presence of the two inhibitors and obtained following data:

Using the entire data set to make a diagnostic plot, answer the following:

 

V0, ?M/sec

[S], ?M

No inhibitor

page3image43240

+ Alice-1 (10 nM)

page3image45016

+ Alice-2 (10 nM)

2

0.062

0.006

page3image50400

0.042

page3image52232

4

0.118

0.012

0.080

8

0.210

0.024

0.157

16

0.348

0.045

0.256

(a) Does Alice-1 or Alice-2 greatly affect the ability of the mutant peptidase to carry out the cleavage reaction (kcat)?

(b) Determine the Km of the mutant proteases for its substrate in the absence and presence of inhibitors.

(c) Which inhibitor is more effective?
(d) What types of inhibitors are Alice-1 and Alice-2?