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Please show work The activity of sphingosine kinase was measured in the presence

ID: 484528 • Letter: P

Question

Please show work

The activity of sphingosine kinase was measured in the presence of threo-dihydrosphingosine, a stereoisomer of sphingosine. The Lineweaver-Burk plot obtained from a kinetic analysis of sphingosine kinase activity carried out in the presence of this inhibitor is shown in Figure 19.6.

1. Calculate the Km and Vmax values in the absence and presence of the inhibitor. What kind of an inhibitor is threo-dihydrosphingosine?

0.2 0.1 0.12 y 0.271x 0.011 0.1 0.08 0.06 0.04 0.0580 x 0.0081 0.02 0.2 0.3 0.1 0.4 ISphingos inel m M 1

Explanation / Answer

In a lineweaver -Burk plor slope = Km/Vmax and intercept = 1/Vmax

In absence of inhibitor :

1/Vmax = 1/0.0081 = 123.46 nmol/min

Km = 0.058 * 123.46 = 7.16 mM

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In presence of inhibitor:

Vax = 1/0.011 =90.91 mmol/min

Km = 0.271 * 90.91 = 24.64 mmol

As both Km and Vmax decreases, this is an uncompititive inhibitor