The Substrate Capture
Mechanism of <i>Mycobacterium
tuberculosis</i> Anthranilate Phosphoribosyltransferase Provides
a Mode for Inhibition
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Abstract
Anthranilate phosphoribosyltransferase (AnPRT, EC 2.4.2.18)
is
a homodimeric enzyme that catalyzes the reaction between 5′-phosphoribosyl
1′-pyrophosphate (PRPP) and anthranilate, as part of the tryptophan
biosynthesis pathway. Here we present the results of the first chemical
screen for inhibitors against <i>Mycobacterium tuberculosis</i> AnPRT (<i>Mtb</i>-AnPRT), along with crystal structures
of <i>Mtb</i>-AnPRT in complex with PRPP and several inhibitors.
Previous work revealed that PRPP is bound at the base of a deep cleft
in <i>Mtb</i>-AnPRT and predicted two anthranilate binding
sites along the tunnel leading to the PRPP binding site. Unexpectedly,
the inhibitors presented here almost exclusively bound at the entrance
of the tunnel, in the presumed noncatalytic anthranilate binding site,
previously hypothesized to have a role in substrate capture. The potencies
of the inhibitors were measured, yielding <i>K</i><sub>i</sub> values of 1.5–119 μM, with the strongest inhibition
displayed by a bianthranilate compound that makes hydrogen bond and
salt bridge contacts with <i>Mtb</i>-AnPRT via its carboxyl
groups. Our results reveal how the substrate capture mechanism of
AnPRT can be exploited to inhibit the enzyme’s activity and
provide a scaffold for the design of improved <i>Mtb</i>-AnPRT inhibitors that may ultimately form the basis of new antituberculosis
drugs with a novel mode of action