Inhibition of chorismate mutase activity of chorismate mutase-prephenate dehydrogenase from Aerobacter aerogenes

Abstract

Inhibition data (I50 values) have been obtained for inhibitors of the chorismate mutase activity of chorismate mutase-prephenate dehydrogenase from Aerobacter aerogenes. Several 1-substituted adamantane derivatives were investigated; the order of decreasing inhibitory activity with the various substituents was -PO32- much greater than -P(OCH3)O2- greater than -CO2- greater than -CH2CO2- greater than -SO2- greater than SO3-. 3-Chloroadamantane-1-acetic acid was slightly less effective than adamantane-1-acetic acid. 2-(1-Carboxy-1,4-dihydrobenzyl)acrylic acid (19), an analogue of prephenate, was an effective inhibitor. Other substances investigated, including 2,4,10-trioxaadamantane-1-acetic acid (15), 5-enolpyruvylshikimic acid (20), and 1-(carboxyethyl)-1,4-dihydrobenzoic acid (18), failed to inhibit chorismate mutase activity under the conditions investigated.

Chorismate mutase-prephenate dehydrogenase from Escherichia coli. 2. Evidence for two different active sites

“…is believed to proceed through a chair-like transition state as depicted in Figure 5. Thus, the approach to the design of inhibitors of the mutase has been to mimic this bicyclic structure (Andrews et al, 1973(Andrews et al, , 1977Chao & Berchtold, 1982; Bartlett & Johnson, 1985;Bartlett et al, 1988). The inhibition constant for the endo-oxabicyclic diacid (Figure 1), synthesized as a racemate, is about 300-fold lower than the Michaelis constant for chorismate in the mutase reaction.…”

“…However, the interaction of adamantane 1-phosphonate at the chorismate binding site was about 5 times greater than at the binding site for prephenate (Table II). The tighter binding of adamantane 1-phosphonate may reflect, in part, its structural similarities to the proposed transition state of the mutase reaction (Andrews et al, 1973(Andrews et al, , 1977Chao & Berchtold, 1982).…”

Characterization of the secreted chorismate mutase from the pathogen Mycobacterium tuberculosis

“…The high K m value at pH 8.7 only allowed the determination of k cat / K m because of the limitation of the maximum chorismate concentration to 1.3 m m . (B) CM transition state analog inhibitors used in this work: 1 [34]; 2 [35]; 3 , adamantane‐1‐phosphonate [38]. (C) Lineweaver–Burk plot [70] with inhibitor 1 .…”

“…Inclusion of 1 m m tryptophan, phenylalanine or tyrosine, or 0.5 m m salicylate in the kinetic assays did not alter CM activity by more than 10% (data not shown). Established CM transition state analog inhibitors, which include the oxabicyclic carboxylic acids 1 [34] and 2 [35,36], and adamantane‐1‐phosphonate 3 [37,38] (Fig. 6B), were tested for their impact on *MtCM activity.…”

“…DNA sequencing was performed on an ABI PRIZM 310 or 3100‐Avant Genetic Analyzer (Applied Biosystems, Foster City, CA, USA) by the chain termination method [55], using the BigDye Terminator Cycle Sequencing Kit from the same company. Chorismate and transition state analogs 1 [34], 2 [35] and 3 [38] (Fig. 6B) were prepared by D Künzler [56], R Pulido [57], A Mandal [36] and S Raillard [37], respectively, following published protocols.…”