Crystal Structure of the Carboxyltransferase Domain of Acetyl-Coenzyme A Carboxylase

Acetyl–coenzyme A carboxylases (ACCs) are required for the biosynthesis and oxidation of long-chain fatty acids. They are targets for therapeutics against obesity and diabetes, and several herbicides function by inhibiting their carboxyltransferase (CT) domain. We determined the crystal structure of the free enzyme and the coenzyme A complex of yeast CT at 2.7 angstrom resolution and found that it comprises two domains, both belonging to the crotonase/ClpP superfamily. The active site is at the interface of a dimer. Mutagenesis and kinetic studies reveal the functional roles of conserved residues here. The herbicides target the active site of CT, providing a lead for inhibitor development against human ACCs.

[1]  J. McGarry,et al.  The mitochondrial carnitine palmitoyltransferase system. From concept to molecular analysis. , 1997, European journal of biochemistry.

[2]  Jimin Wang,et al.  The Structure of ClpP at 2.3 Å Resolution Suggests a Model for ATP-Dependent Proteolysis , 1997, Cell.

[3]  R. Haselkorn,et al.  The Carboxyltransferase Activity of the Apicoplast Acetyl-CoA Carboxylase of Toxoplasma gondii Is the Target of Aryloxyphenoxypropionate Inhibitors* , 2002, The Journal of Biological Chemistry.

[4]  J. Hiltunen,et al.  Crystal structure of enoyl‐coenzyme A (CoA) hydratase at 2.5 angstroms resolution: a spiral fold defines the CoA‐binding pocket. , 1996, The EMBO journal.

[5]  F. R. van der Leij,et al.  Molecular enzymology of carnitine transfer and transport. , 2001, Biochimica et biophysica acta.

[6]  Grover L Waldrop,et al.  A Biotin Analog Inhibits Acetyl-CoA Carboxylase Activity and Adipogenesis* , 2002, The Journal of Biological Chemistry.

[7]  K. Sharp,et al.  Protein folding and association: Insights from the interfacial and thermodynamic properties of hydrocarbons , 1991, Proteins.

[8]  Martin M. Matzuk,et al.  Continuous Fatty Acid Oxidation and Reduced Fat Storage in Mice Lacking Acetyl-CoA Carboxylase 2 , 2001, Science.

[9]  S. Wakil,et al.  Fatty acid synthesis and its regulation. , 1983, Annual review of biochemistry.

[10]  J. Lenhard,et al.  Preclinical developments in type 2 diabetes. , 2002, Advanced drug delivery reviews.

[11]  M. Lane,et al.  Acetyl coenzyme A carboxylase system of Escherichia coli. Studies on the mechanisms of the biotin carboxylase- and carboxyltransferase-catalyzed reactions. , 1974, The Journal of biological chemistry.

[12]  D. Dunaway-Mariano,et al.  Structure of 4-chlorobenzoyl coenzyme A dehalogenase determined to 1.8 A resolution: an enzyme catalyst generated via adaptive mutation. , 1996, Biochemistry.

[13]  Mike Carson,et al.  Ribbon models of macromolecules , 1987 .

[14]  J. W. Campbell,et al.  Bacterial fatty acid biosynthesis: targets for antibacterial drug discovery. , 2001, Annual review of microbiology.

[15]  J. Hiltunen,et al.  THE CRYSTAL STRUCTURE , 2010 .

[16]  H M Holden,et al.  New reactions in the crotonase superfamily: structure of methylmalonyl CoA decarboxylase from Escherichia coli. , 2000, Biochemistry.

[17]  R. Haselkorn,et al.  An isoleucine/leucine residue in the carboxyltransferase domain of acetyl-CoA carboxylase is critical for interaction with aryloxyphenoxypropionate and cyclohexanedione inhibitors , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[18]  W. Hendrickson.  Determination of macromolecular structures from anomalous diffraction of synchrotron radiation. , 1991, Science.

[19]  Grover L Waldrop,et al.  Multi-subunit acetyl-CoA carboxylases. , 2002, Progress in lipid research.

[20]  B. Matthews,et al.  Competing protein:protein interactions are proposed to control the biological switch of the E coli biotin repressor , 2001, Protein science : a publication of the Protein Society.

[21]  J. Knowles.  The mechanism of biotin-dependent enzymes. , 1989, Annual review of biochemistry.