3GGS

Human purine nucleoside phosphorylase double mutant E201Q,N243D complexed with 2-fluoro-2'-deoxyadenosine

3GGS の概要

• エントリーDOI: 10.2210/pdb3ggs/pdb
• 関連するPDBエントリー: 3GB9
• 分子名称: Purine nucleoside phosphorylase, 5-(6-AMINO-2-FLUORO-PURIN-9-YL)-2-HYDROXYMETHYL-TETRAHYDRO-FURAN-3-OL, SULFATE ION, ... (4 entities in total)
• 機能のキーワード: enzyme-prodrug complex, disease mutation, glycosyltransferase, transferase
• 由来する生物種: Homo sapiens (Human)
• 細胞内の位置: Cytoplasm, cytoskeleton : P00491
• タンパク質・核酸の鎖数: 3
• 化学式量合計: 107159.56

構造登録者

Sawaya, M.R.,Afshar, S. (登録日: 2009-03-02, 公開日: 2009-04-14, 最終更新日: 2023-09-06)

主引用文献

Afshar, S.,Sawaya, M.R.,Morrison, S.L.
Structure of a mutant human purine nucleoside phosphorylase with the prodrug, 2-fluoro-2'-deoxyadenosine and the cytotoxic drug, 2-fluoroadenine.
Protein Sci., 18:1107-1114, 2009

PubMed Abstract: A double mutant of human purine nucleoside phosphorylase (hDM) with the amino acid mutations Glu201Gln:Asn243Asp cleaves adenosine-based prodrugs to their corresponding cytotoxic drugs. When fused to an anti-tumor targeting component, hDM is targeted to tumor cells, where it effectively catalyzes phosphorolysis of the prodrug, 2-fluoro-2'-deoxyadenosine (F-dAdo) to the cytotoxic drug, 2-fluoroadenine (F-Ade). This cytotoxicity should be restricted only to the tumor microenvironment, because the endogenously expressed wild type enzyme cannot use adenosine-based prodrugs as substrates. To gain insight into the interaction of hDM with F-dAdo, we have determined the crystal structures of hDM with F-dAdo and F-Ade. The structures reveal that despite the two mutations, the overall fold of hDM is nearly identical to the wild type enzyme. Importantly, the residues Gln201 and Asp243 introduced by the mutation form hydrogen bond contacts with F-dAdo that result in its binding and catalysis. Comparison of substrate and product complexes suggest that the side chains of Gln201 and Asp243 as well as the purine base rotate during catalysis possibly facilitating cleavage of the glycosidic bond. The two structures suggest why hDM, unlike the wild-type enzyme, can utilize F-dAdo as substrate. More importantly, they provide a critical foundation for further optimization of cleavage of adenosine-based prodrugs, such as F-dAdo by mutants of human purine nucleoside phosphorylase.

PubMed: 19388075
DOI: 10.1002/pro.91

実験手法

X-RAY DIFFRACTION (2.52 Å)