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Acyl-CoA dehydrogenases and acyl-CoA oxidases are two closely related FAD-containing enzyme families that are present inmitochondriaandperoxisomes, respectively. They catalyze the dehydrogenation of acyl-CoA thioesters to the correspondingtrans-2-enoyl-CoA. This review examines the structure of medium chain acyl-CoA dehydrogenase, as a representative of the dehydrogenase family, with respect to the catalytic mechanism and its broad chain length specificity. Comparing the structures of four other acyl-CoA dehydrogenases provides further insights into the structural basis for the substrate specificityof eachof these enzymes. . | Eur. J. Biochem. 271 483-493 2004 FEBS 2004 doi 10.1046 j.1432-1033.2003.03948.x MINIREVIEW Acyl-CoA dehydrogenases and acyl-CoA oxidases Structural basis for mechanistic similarities and differences Jung-Ja P. Kim1 and Retsu Miura2 1 Department of Biochemistry Medical College of Wisconsin Milwaukee WI USA department of Molecular Enzymology Graduate School of Medical Sciences Kumamoto University Japan Acyl-CoA dehydrogenases and acyl-CoA oxidases are two closely related FAD-containing enzyme families that are present in mitochondria and peroxisomes respectively. They catalyze the dehydrogenation of acyl-CoA thioesters to the corresponding trans-2-enoyl-CoA. This review examines the structure of medium chain acyl-CoA dehydrogenase as a representative of the dehydrogenase family with respect to the catalytic mechanism and its broad chain length specificity. Comparing the structures of four other acyl-CoA dehydrogenases provides further insights into the structural basis for the substrate specificity of each of these enzymes. In addition the structure of peroxisomal acyl-CoA oxidase II from rat liver is compared to that of medium chain acyl-CoA dehydrogenase and the structural basis for their different oxidative half reactions is discussed. Introduction There are nine known members in the acyl-CoA dehydrogenase ACAD family including the recently identified ACAD-9 which is homologous to the very long chain acyl-CoA dehydrogenase 1 . Five members of the ACAD family are involved in fatty acid b-oxidation these are short medium long and very long chain acyl-CoA dehydrogenase SCAD mCaD lCaD and VLCAD1 resepctively and ACAD-9 hereafter referred to as VLCAD2 . The four other members are involved in amino acid oxidation pathways they are iso 3 valeryl-CoA dehydrogenase i3VD for leucine iso 2 valeryl-CoA dehydrogenase i2VD also known as short branched chain acyl-CoA dehydrogenase or 2-methylbutyryl-CoA dehydrogenase for isoleucine isobutyryl-CoA dehydrogenase iBD for valine .