3TE4

Crystal structure of dopamine N Acetyltransferase in complex with acetyl-COA from Drosophila Melanogaster

3TE4 の概要

• エントリーDOI: 10.2210/pdb3te4/pdb
• 分子名称: Dopamine N acetyltransferase, isoform A, ACETYL COENZYME *A (3 entities in total)
• 機能のキーワード: transferase, dopamine/acetyl coa, n-acetyltransferase domain
• 由来する生物種: Drosophila melanogaster (Fruit fly)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 25251.62

構造登録者

Cheng, K.C.,Huang, S.H.,Lyu, P.C. (登録日: 2011-08-12, 公開日: 2012-07-11, 最終更新日: 2024-03-20)

主引用文献

Cheng, K.C.,Liao, J.N.,Lyu, P.C.
Crystal structure of the dopamine N-acetyltransferase-acetyl-CoA complex provides insights into the catalytic mechanism
Biochem.J., 446:395-404, 2012

PubMed Abstract: The daily cycle of melatonin biosynthesis in mammals is regulated by AANAT (arylalkylamine N-acetyltransferase; EC 2.3.1.87), making it an attractive target for therapeutic control of abnormal melatonin production in mood and sleep disorders. Drosophila melanogaster Dat (dopamine N-acetyltransferase) is an AANAT. Until the present study, no insect Dat structure had been solved, and, consequently, the structural basis for its acetyl-transfer activity was not well understood. We report in the present paper the high-resolution crystal structure for a D. melanogaster Dat-AcCoA (acetyl-CoA) complex obtained using one-edge (selenium) single-wavelength anomalous diffraction. A binding study using isothermal titration calorimetry suggested that the cofactor bound to Dat first before substrate. Examination of the complex structure and a substrate-docked model indicated that Dat contains a novel AANAT catalytic triad. Site-directed mutagenesis, kinetic studies and pH-rate profiles confirmed that Glu(47), Ser(182) and Ser(186) were critical for catalysis. Collectively, the results of the present study suggest that Dat possesses a specialized active site structure dedicated to a catalytic mechanism.

PubMed: 22716280
DOI: 10.1042/BJ20120520

実験手法

X-RAY DIFFRACTION (1.46 Å)