4FE3

Structure of murine cytosolic 5'-nucleotidase III complexed with uridinine monophosphate

4FE3 の概要

• エントリーDOI: 10.2210/pdb4fe3/pdb
• 関連するPDBエントリー: 2G06 2G09
• 分子名称: Cytosolic 5'-nucleotidase 3, BETA-MERCAPTOETHANOL, SODIUM ION, ... (6 entities in total)
• 機能のキーワード: substrate complex, had-like, nucleotidase, protein binding
• 由来する生物種: Mus musculus (mouse)
• 細胞内の位置: Cytoplasm (Potential): Q9D020
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 34359.64

構造登録者

Bitto, E.,Bingman, C.A. (登録日: 2012-05-29, 公開日: 2012-08-29, 最終更新日: 2023-09-13)

主引用文献

Grobosky, C.L.,Lopez, J.B.,Rennie, S.,Skopelitis, D.J.,Wiest, A.T.,Bingman, C.A.,Bitto, E.
Structural Basis of Substrate Specificity and Selectivity of Murine Cytosolic 5'-Nucleotidase III.
J.Mol.Biol., 423:540-554, 2012

PubMed Abstract: Cytosolic 5'-nucleotidase III (cN-III) is responsible for selective degradation of pyrimidine 5'-monoribonucleotides during maturation of reticulocytes to erythrocytes. The lack of this enzymatic activity due to genetic aberrations or lead poisoning results in a mild to moderate nonspherocytic hemolytic anemia. In affected individuals, pyrimidine nucleotides as well as their precursor polymers and their off-path metabolites accumulate in erythrocytes, interfering with their proper function in ways that are not yet fully understood. This report describes the first X-ray structure of a catalytically inactivated variant of murine cN-III with a natural substrate, uridine 5'-monophosphate, in the active site at 1.74Å resolution. The structure captures in an atomic detail the closed conformation that cN-III adopts upon substrate binding. Structure and sequence analysis coupled with enzymatic characterization of several mutants confirmed that the aromatic ring of a nitrogenous base of substrate nucleotide is stabilized by parallel π-stacking interactions with conserved aromatic rings of Trp113 and His68. The nitrogenous base is further stabilized by T-shaped stacking with the conserved aromatic ring of Tyr114, as well as by polar contacts with side chains of Thr66 and Ser117. Two water molecules help to stabilize the nucleotide binding by bridging it to protein residues Asp72 and His68 via hydrogen bonds. Finally, fully conserved Glu96 is responsible for recognition of ribose ring via two hydrogen bonds. The presented substrate complex structure elucidates how cN-III achieves specificity for pyrimidine 5'-nucleotides and how it selects against purine 5'-nucleotides.

PubMed: 22925580
DOI: 10.1016/j.jmb.2012.08.014

実験手法

X-RAY DIFFRACTION (1.74 Å)