3OCW

Structure of Recombinant Haemophilus influenzae e(P4) Acid Phosphatase mutant D66N complexed with 3'-AMP

3OCW の概要

• エントリーDOI: 10.2210/pdb3ocw/pdb
• 関連するPDBエントリー: 3OCU 3OCV 3OCX 3OCY 3OCZ 3et4
• 分子名称: Lipoprotein E, MAGNESIUM ION, [(2R,3S,4R,5R)-5-(6-aminopurin-9-yl)-4-hydroxy-2-(hydroxymethyl)oxolan-3-yl] dihydrogen phosphate, ... (4 entities in total)
• 機能のキーワード: hydrolase, outer membrane
• 由来する生物種: Haemophilus influenzae
• 細胞内の位置: Cell outer membrane; Lipid-anchor: P26093
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 29991.52

構造登録者

Singh, H.,Schuermann, J.,Reilly, T.,Calcutt, M.,Tanner, J. (登録日: 2010-08-10, 公開日: 2010-10-20, 最終更新日: 2023-09-06)

主引用文献

Singh, H.,Schuermann, J.P.,Reilly, T.J.,Calcutt, M.J.,Tanner, J.J.
Recognition of nucleoside monophosphate substrates by Haemophilus influenzae class C acid phosphatase.
J.Mol.Biol., 404:639-649, 2010

PubMed Abstract: The e (P4) phosphatase from Haemophilus influenzae functions in a vestigial NAD(+) utilization pathway by dephosphorylating nicotinamide mononucleotide to nicotinamide riboside. P4 is also the prototype of class C acid phosphatases (CCAPs), which are nonspecific 5',3'-nucleotidases localized to the bacterial outer membrane. To understand substrate recognition by P4 and other class C phosphatases, we have determined the crystal structures of a substrate-trapping mutant P4 enzyme complexed with nicotinamide mononucleotide, 5'-AMP, 3'-AMP, and 2'-AMP. The structures reveal an anchor-shaped substrate-binding cavity comprising a conserved hydrophobic box that clamps the nucleotide base, a buried phosphoryl binding site, and three solvent-filled pockets that contact the ribose and the hydrogen-bonding edge of the base. The span between the hydrophobic box and the phosphoryl site is optimal for recognizing nucleoside monophosphates, explaining the general preference for this class of substrate. The base makes no hydrogen bonds with the enzyme, consistent with an observed lack of base specificity. Two solvent-filled pockets flanking the ribose are key to the dual recognition of 5'-nucleotides and 3'-nucleotides. These pockets minimize the enzyme's direct interactions with the ribose and provide sufficient space to accommodate 5' substrates in an anti conformation and 3' substrates in a syn conformation. Finally, the structures suggest that class B acid phosphatases and CCAPs share a common strategy for nucleotide recognition.

PubMed: 20934434
DOI: 10.1016/j.jmb.2010.09.065

実験手法

X-RAY DIFFRACTION (1.85 Å)