2EWB

The crystal structure of Bovine Lens Leucine Aminopeptidase in complex with zofenoprilat

2EWB の概要

• エントリーDOI: 10.2210/pdb2ewb/pdb
• 関連するPDBエントリー: 1LAM
• 分子名称: Cytosol aminopeptidase, ZINC ION, SODIUM ION, ... (7 entities in total)
• 機能のキーワード: aminopeptidase, metallopeptidase, hydrolase
• 由来する生物種: Bos taurus (cattle)
• 細胞内の位置: Cytoplasm: P00727
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 53882.98

構造登録者

Alterio, V.,Pedone, C.,De Simone, G. (登録日: 2005-11-02, 公開日: 2006-04-04, 最終更新日: 2023-08-23)

主引用文献

Cappiello, M.,Alterio, V.,Amodeo, P.,Del Corso, A.,Scaloni, A.,Pedone, C.,Moschini, R.,De Donatis, G.M.,De Simone, G.,Mura, U.
Metal Ion Substitution in the Catalytic Site Greatly Affects the Binding of Sulfhydryl-Containing Compounds to Leucyl Aminopeptidase.
Biochemistry, 45:3226-3234, 2006

PubMed Abstract: Bovine lens leucyl aminopeptidase (blLAP), a homohexameric metallopeptidase preferring bulky and hydrophobic amino acids at the N-terminus of (di)peptides, contains two Zn(2+) ions per subunit that are essential for catalytic activity. They may be replaced by other divalent cations with different exchange kinetics. The protein readily exchangeable site (site 1) can be occupied by Zn(2+), Mn(2+), Mg(2+), or Co(2+), while the tight binding site (site 2) can be occupied by Zn(2+) or Co(2+). We recently reported that introduction of Mn(2+) into site 1 generates a novel activity of blLAP toward CysGly [Cappiello, M., et al. (2004) Biochem. J. 378, 35-44], which in contrast is not hydrolyzed by the (Zn/Zn) enzyme. This finding, while disclosing a potential specific role for blLAP in glutathione metabolism, raised a question about the features required for molecules to be a substrate for the enzyme. To clarify the interaction of the enzyme with sulfhydryl-containing derivatives, (Zn/Zn)- and (Mn/Zn)blLAP forms were prepared and functional-structural studies were undertaken. Thus, a kinetic analysis of various compounds with both enzyme forms was performed; the crystal structure of (Zn/Zn)blLAP in complex with the peptidomimetic derivative Zofenoprilat was determined, and a modeling study on the CysGly-(Zn/Zn)blLAP complex was carried out. This combined approach provided insight into the interaction of blLAP with sulfhydryl-containing derivatives, showing that the metal exchange in site 1 modulates binding to these molecules that may result in enzyme substrates or inhibitors, depending on the nature of the metal.

PubMed: 16519517
DOI: 10.1021/bi052069v

実験手法

X-RAY DIFFRACTION (1.85 Å)