crystal structure and substrate specificity of drosophila 3,4-dihydroxyphenylalanine decarboxylase晶体结构和底物特异性的果蝇,4-dihydroxyphenylalanine脱羧酶.pdfVIP

Crystal Structure and Substrate Specificity of Drosophila 3,4-Dihydroxyphenylalanine Decarboxylase 1 1 2 3 1 Qian Han , Haizhen Ding , Howard Robinson , Bruce M. Christensen , Jianyong Li * 1 Department of Biochemistry, Virginia Tech, Blacksburg, Virginia, United States of America, 2 Biology Department, Brookhaven National Laboratory, Upton, New York, United States of America, 3 Department of Pathobiological Sciences, University of Wisconsin, Madison, Wisconsin, United States of America Abstract Background: 3,4-Dihydroxyphenylalanine decarboxylase (DDC), also known as aromatic L-amino acid decarboxylase, catalyzes the decarboxylation of a number of aromatic L-amino acids. Physiologically, DDC is responsible for the production of dopamine and serotonin through the decarboxylation of 3,4-dihydroxyphenylalanine and 5-hydroxy- tryptophan, respectively. In insects, both dopamine and serotonin serve as classical neurotransmitters, neuromodu- lators, or neurohormones, and dopamine is also involved in insect cuticle formation, eggshell hardening, and immune responses. Principal Findings: In this study, we expressed a typical DDC enzyme from Drosophila melanogaster, critically analyzed its substrate specificity and biochemical properties, determined its crystal structure at 1.75 Angstrom resolution, and evaluated the roles residues T82 and H192 play in substrate binding and enzyme catalysis through site-directed mutagenesis of the enzyme. Our results establish that this DDC functions exclusively on the production of dopamine and serotonin, with no activity to tyrosine or tryptophan and catalyzes the formation of serotonin more efficiently than dopamine. Conclusions: The crystal structure of Drosophil