3UZB

Crystal Structures of Branched-Chain Aminotransferase from Deinococcus radiodurans Complexes with alpha-Ketoisocaproate and L-Glutamate Suggest Its Radio-Resistance for Catalysis

3UZB の概要

• エントリーDOI: 10.2210/pdb3uzb/pdb
• 関連するPDBエントリー: 3UYY 3UZO
• 分子名称: Branched-chain-amino-acid aminotransferase, PYRIDOXAL-5'-PHOSPHATE, 2-OXO-4-METHYLPENTANOIC ACID, ... (4 entities in total)
• 機能のキーワード: bcat, amino-acid biosynthesis, aminotransferase, branched-chain amino acid biosynthesis, pyridoxal phosphate, transferase, alpha-ketoisocaproate
• 由来する生物種: Deinococcus radiodurans
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 159370.43

構造登録者

Chen, C.D.,Huang, Y.C.,Chuankhayan, P.,Hsieh, Y.C.,Huang, T.F.,Lin, C.H.,Guan, H.H.,Liu, M.Y.,Chang, W.C.,Chen, C.J. (登録日: 2011-12-07, 公開日: 2012-12-12, 最終更新日: 2023-11-08)

主引用文献

Chen, C.D.,Lin, C.H.,Chuankhayan, P.,Huang, Y.C.,Hsieh, Y.C.,Huang, T.F.,Guan, H.H.,Liu, M.Y.,Chang, W.C.,Chen, C.J.
Crystal Structures of Complexes of the Branched-Chain Aminotransferase from Deinococcus radiodurans with alpha-Ketoisocaproate and L-Glutamate Suggest the Radiation Resistance of This Enzyme for Catalysis
J.Bacteriol., 194:6206-6216, 2012

PubMed Abstract: Branched-chain aminotransferases (BCAT), which utilize pyridoxal 5'-phosphate (PLP) as a cofactor, reversibly catalyze the transfer of the α-amino groups of three of the most hydrophobic branched-chain amino acids (BCAA), leucine, isoleucine, and valine, to α-ketoglutarate to form the respective branched-chain α-keto acids and glutamate. The BCAT from Deinococcus radiodurans (DrBCAT), an extremophile, was cloned and expressed in Escherichia coli for structure and functional studies. The crystal structures of the native DrBCAT with PLP and its complexes with L-glutamate and α-ketoisocaproate (KIC), respectively, have been determined. The DrBCAT monomer, comprising 358 amino acids, contains large and small domains connected with an interdomain loop. The cofactor PLP is located at the bottom of the active site pocket between two domains and near the dimer interface. The substrate (L-glutamate or KIC) is bound with key residues through interactions of the hydrogen bond and the salt bridge near PLP inside the active site pocket. Mutations of some interaction residues, such as Tyr71, Arg145, and Lys202, result in loss of the specific activity of the enzymes. In the interdomain loop, a dynamic loop (Gly173 to Gly179) clearly exhibits open and close conformations in structures of DrBCAT without and with substrates, respectively. DrBCAT shows the highest specific activity both in nature and under ionizing radiation, but with lower thermal stability above 60 °C, than either BCAT from Escherichia coli (eBCAT) or from Thermus thermophilus (HB8BCAT). The dimeric molecular packing and the distribution of cysteine residues at the active site and the molecular surface might explain the resistance to radiation but small thermal stability of DrBCAT.

PubMed: 22984263
DOI: 10.1128/JB.01659-12

実験手法

X-RAY DIFFRACTION (3 Å)