1L9E

Role of Histidine 269 in Catalysis by Monomeric Sarcosine Oxidase

1L9E の概要

• エントリーDOI: 10.2210/pdb1l9e/pdb
• 関連するPDBエントリー: 1L9C 1L9D 1L9F
• 分子名称: Monomeric sarcosine oxidase, CHLORIDE ION, FLAVIN-ADENINE DINUCLEOTIDE, ... (5 entities in total)
• 機能のキーワード: flavoprotein, oxidase, oxidoreductase
• 由来する生物種: Bacillus sp.
• 細胞内の位置: Cytoplasm: P40859
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 87982.89

構造登録者

Zhao, G.,Song, H.,Chen, Z.-w.,Mathews, F.S.,Jorns, M.S. (登録日: 2002-03-22, 公開日: 2002-08-30, 最終更新日: 2024-10-16)

主引用文献

Zhao, G.,Song, H.,Chen, Z.W.,Mathews, F.S.,Jorns, M.S.
Monomeric sarcosine oxidase: role of histidine 269 in catalysis.
Biochemistry, 41:9751-9764, 2002

PubMed Abstract: Conservative mutation of His269 (to Asn, Ala, or Gln) does not-significantly affect the expression of monomeric sarcosine oxidase (MSOX), covalent flavinylation, the physicochemical properties of bound FAD, or the overall protein structure. Turnover with sarcosine and the limiting rate of the reductive half-reaction with L-proline at pH 8.0 are, however, nearly 2 orders of magnitude slower than that with with wild-type MSOX. The crystal structure of the His269Asn complex with pyrrole-2-carboxylate shows that the pyrrole ring of the inhibitor is displaced as compared with wild-type MSOX. The His269 mutants all form charge-transfer complexes with pyrrole-2-carboxylate or methylthioacetate, but the charge-transfer bands are shifted to shorter wavelengths (higher energy) as compared with wild-type MSOX. Both wild-type MSOX and the His269Asn mutant bind the zwitterionic form of L-proline. The E(ox).L-proline complex formed with the His269Asn mutant or wild-type MSOX contains an ionizable group (pK(a) = 8.0) that is required for conversion of the zwitterionic L-proline to the reactive anionic form, indicating that His269 is not the active-site base. We propose that the change in ligand orientation observed upon mutation of His269 results in a less than optimal overlap of the highest occupied orbital of the ligand with the lowest unoccupied orbital of the flavin. The postulated effect on orbital overlap may account for the increased energy of charge-transfer bands and the slower rates of electron transfer observed for mutant enzyme complexes with charge-transfer ligands and substrates, respectively.

PubMed: 12146941
DOI: 10.1021/bi020286f

実験手法

X-RAY DIFFRACTION (1.85 Å)