2A1H

X-ray crystal structure of human mitochondrial branched chain aminotransferase (BCATm) complexed with gabapentin

2A1H の概要

• エントリーDOI: 10.2210/pdb2a1h/pdb
• 関連するPDBエントリー: 1EKF 1EKP 1EKV 1KT8 1KTA
• 分子名称: branched chain aminotransferase, PYRIDOXAL-5'-PHOSPHATE, [1-(AMINOMETHYL)CYCLOHEXYL]ACETIC ACID, ... (5 entities in total)
• 機能のキーワード: fold type iv, transferase
• 由来する生物種: Homo sapiens (human)
• 細胞内の位置: Isoform A: Mitochondrion. Isoform B: Cytoplasm: O15382
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 83632.84

構造登録者

Goto, M.,Miyahara, I.,Hirotsu, K.,Conway, M.,Yennawar, N.,Islam, M.M.,Hutson, S.M. (登録日: 2005-06-20, 公開日: 2005-09-06, 最終更新日: 2025-03-26)

主引用文献

Goto, M.,Miyahara, I.,Hirotsu, K.,Conway, M.,Yennawar, N.,Islam, M.M.,Hutson, S.M.
Structural determinants for branched-chain aminotransferase isozyme-specific inhibition by the anticonvulsant drug gabapentin.
J.Biol.Chem., 280:37246-37256, 2005

PubMed Abstract: This study presents the first three-dimensional structures of human cytosolic branched-chain aminotransferase (hBCATc) isozyme complexed with the neuroactive drug gabapentin, the hBCATc Michaelis complex with the substrate analog, 4-methylvalerate, and the mitochondrial isozyme (hBCATm) complexed with gabapentin. The branched-chain aminotransferases (BCAT) reversibly catalyze transamination of the essential branched-chain amino acids (leucine, isoleucine, valine) to alpha-ketoglutarate to form the respective branched-chain alpha-keto acids and glutamate. The cytosolic isozyme is the predominant BCAT found in the nervous system, and only hBCATc is inhibited by gabapentin. Pre-steady state kinetics show that 1.3 mm gabapentin can completely inhibit the binding of leucine to reduced hBCATc, whereas 65.4 mm gabapentin is required to inhibit leucine binding to hBCATm. Structural analysis shows that the bulky gabapentin is enclosed in the active-site cavity by the shift of a flexible loop that enlarges the active-site cavity. The specificity of gabapentin for the cytosolic isozyme is ascribed at least in part to the location of the interdomain loop and the relative orientation between the small and large domain which is different from these relationships in the mitochondrial isozyme. Both isozymes contain a CXXC center and form a disulfide bond under oxidizing conditions. The structure of reduced hBCATc was obtained by soaking the oxidized hBCATc crystals with dithiothreitol. The close similarity in active-site structures between cytosolic enzyme complexes in the oxidized and reduced states is consistent with the small effect of oxidation on pre-steady state kinetics of the hBCATc first half-reaction. However, these kinetic data do not explain the inactivation of hBCATm by oxidation of the CXXC center. The structural data suggest that there is a larger effect of oxidation on the interdomain loop and residues surrounding the CXXC center in hBCATm than in hBCATc.

PubMed: 16141215
DOI: 10.1074/jbc.M506486200

実験手法

X-RAY DIFFRACTION (1.8 Å)