Crystal Structure of the Carboxyltransferase Domain of Acetyl-Coenzyme A Carboxylase in Complex with CP-640186

Abstract

Acetyl-coenzyme A carboxylases (ACCs) are important targets for the development of therapeutic agents against obesity, diabetes, and other diseases. CP-640186 is a potent inhibitor of mammalian ACCs and can reduce body weight and improve insulin sensitivity in test animals. It is believed to target the carboxyltransferase (CT) domain of these enzymes. Here we report the crystal structure of the yeast CT domain in complex with CP-640186. The inhibitor is bound in the active site at the interface of a dimer of the CT domain. CP-640186 has tight interactions with the putative biotin binding site in the CT domain and demonstrates a distinct mode of inhibiting the CT activity as compared to the herbicides that inhibit plant ACCs. The affinity of inhibitors for the CT domain has been assessed using kinetic and fluorescence anisotropy binding studies. The structural information identifies three regions for drug binding in the active site of CT.

The enzymes of biotin dependent CO2 metabolism: What structures reveal about their reaction mechanisms

“…Subsequent structural analysis of a complex between the yeast carboxyltransferase domain and CP-640186 showed that the inhibitor binds at the interface of the carboxyltransferase dimer, and it was concluded that this is also the biotin-binding site. 85 It should be noted, however, that a kinetic analysis of the inhibitor binding versus biotin was not reported, and thus the proposed biotinbinding site is still open to discussion.…”

The cyclic keto-enol insecticide spirotetramat inhibits insect and spider mite acetyl-CoA carboxylases by interfering with the carboxyltransferase partial reaction

“…The steady-state kinetic data were interpreted as distinct binding sites for the known ACC inhibitor classes within the CT domain, a concept that was later substantiated by X-ray structural analysis (Harwood et al, 2003, Zhang et al, 2004. In the context of this discussion, the binding mode of SPT-enol may be similar, but not identical to the aryloxyphenoxypropionates and cyclohexanediones.…”

“…Finally, a novel class of bipiperidylcarboxamide CT domain inhibitors of mammalian ACCs was discovered by highthroughput biochemical screening (Corbett et al, 2010). Interestingly, X-ray structural data demonstrated different binding positions within the CT domain for the bispiperidylcarboxamide CP-640186 and the herbicide haloxyfop (Zhang et al, 2004).…”

“…In the mite protein, a leucine is present at this position, and we introduced the L1736I and L1736A substitutions to assess the effect on mite enzyme catalytic activity and inhibitor sensitivity (Figure 7). The second position is A1739 (corresponding to the conserved S1783 in monocotyledonous and dicotyledonous plants) which is located in the deep binding pocket (Zhang et al, 2004). The substitutions A1739S and A1739V were introduced (Fig.…”

Modeling the coverage of an AFM tip by enzymes and its application in nanobiosensors

“…In this study, a coarse grained model is proposed to determine the interaction between acetyl-CoA carboxylase (ACC) and diclofop, a commercial herbicide known to block ACC activity by occupying its binding pocket [13][14][15][16][17] in a non-covalent, reversible interaction. The coarse-grained model for the AFM tip surface covered by enzymes uses atomistic information obtained from MD simulations and technical specifications from the manufacturer of the AFM tip.…”