Based on your Lineweaver-Burk plot, what is the mode of PaESBL-1 inhibition by NSAMB?

Biochemistry
9th Edition
ISBN:9781319114671
Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Publisher:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Chapter1: Biochemistry: An Evolving Science
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Based on your Lineweaver-Burk plot, what is the mode of PaESBL-1 inhibition by NSAMB?

What is the equation for the double-reciprocal plot at 20 nMnM NAMB?

You contact an academic lab that specializes in synthesizing boronic acid analogs that have been shown to act as reversible inhibitors of the AmpC B-lactamase from Escherichia coll. The lab agrees to send you several of its compounds to test against PAESBL-1. You begin by
looking at the potential of (napthylmethanesulfonylamino)methaneboronic acid (NSAMB) to inhibit the P. aeruginosa B-lactamase:
он
B
O=S- NH
OH
NSAMB
You perform a classic reversible inhibition experiment, where you measure the initial velocity for a range of substrate and inhibitor concentrations. For this assay, you use the artificial chromogenic substrate, CENTA:
`NO
CENTA
CENTA is hydrolyzed by B-lactamases to yield a product with absorbance at 405 nm.
The results of your experiment are listed in the table:
Initial velocity measurements (mAbs/s)
CENTA concentration
(µM)
[NSAMB) = 0 nM
[NSAMB) = 20 nM
[NSAMB] = 40 nM
[NSAMB) = 80 nM
15
0.125
0.102
0.087
0.066
30
0.167
0.145
0.129
0.105
60
0.200
0.184
0.170
0.148
120
0.222
0.212
0.202
0.186
240
0.235
0.229
0.224
0.213
Transcribed Image Text:You contact an academic lab that specializes in synthesizing boronic acid analogs that have been shown to act as reversible inhibitors of the AmpC B-lactamase from Escherichia coll. The lab agrees to send you several of its compounds to test against PAESBL-1. You begin by looking at the potential of (napthylmethanesulfonylamino)methaneboronic acid (NSAMB) to inhibit the P. aeruginosa B-lactamase: он B O=S- NH OH NSAMB You perform a classic reversible inhibition experiment, where you measure the initial velocity for a range of substrate and inhibitor concentrations. For this assay, you use the artificial chromogenic substrate, CENTA: `NO CENTA CENTA is hydrolyzed by B-lactamases to yield a product with absorbance at 405 nm. The results of your experiment are listed in the table: Initial velocity measurements (mAbs/s) CENTA concentration (µM) [NSAMB) = 0 nM [NSAMB) = 20 nM [NSAMB] = 40 nM [NSAMB) = 80 nM 15 0.125 0.102 0.087 0.066 30 0.167 0.145 0.129 0.105 60 0.200 0.184 0.170 0.148 120 0.222 0.212 0.202 0.186 240 0.235 0.229 0.224 0.213
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