Burkweaver line plot

Ronanc1

Hey guys i need some help with my biochemistry I need to know how to construct a burkweaver line plot

Ive run an assay say with alkaline phosphotase with known standards etc and say i get readings like so from the Spectrometer (im just making up some no.s) So i plot them on a standard curve

1 0.000
2 .208
3 .400
4 .600
5 .800
6 .900

I then run the assay again with an inhibitor and get another six absorbances say
1 a
2 b
3 c
4 d
5 e
6 f

How do i construct a lineweaver burk plot what goes where how do i get the points etc all i know is whats going on the axis' 1/V0 and 1/

Any help appreciated guys

Improbable

Ronanc1 wrote: »

Hey guys i need some help with my biochemistry I need to know how to construct a burkweaver line plot

Ive run an assay say with alkaline phosphotase with known standards etc and say i get readings like so from the Spectrometer (im just making up some no.s) So i plot them on a standard curve

1 0.000
2 .208
3 .400
4 .600
5 .800
6 .900

I then run the assay again with an inhibitor and get another six absorbances say
1 a
2 b
3 c
4 d
5 e
6 f

How do i construct a lineweaver burk plot what goes where how do i get the points etc all i know is whats going on the axis' 1/V0 and 1/

Any help appreciated guys

Given that you have an uninhibited reaction and an inhibited reaction, I'm going to assume that the point of doing the Lineweaver-Burk plot is to get the type of inhibition?

1/v goes on the Y axis where v is the rate of reaction.
1/ goes on the X axis where is the substrate concentration.

Because you ran the experiment with known standards (where you already know the concentration), and the experimental results will give you v, the rate of reaction, you just plot 1/v against 1/.

You could get a straight line.

The point at which the line intercepts the Y axis is equal to 1/Vmax
You can then extend the line back to the X axis intercept.
The point at which the line meets the X axis is equal to -1/Km

Also, the slope of the line is equal to Km/Vmax

Do both plots on the same sheet of graph paper. If the 2 lines have the same Y axis intercept but different X axis intercept, it is a competitive inhibitor. If they have the same X axis intercept but different Y axis intercept, then it is a non-competitive inhibitor.

Ronanc1

Thanks improbable yeah i should have mentioned the inhibition type all that info was really helpful thanks so much:D