PDBsum entry 2vq7

Oxidoreductase

PDB id: 2vq7

Contents

Protein chains

443 a.a. *

Ligands

FAD ×4

NAP ×4

EPE ×4

PGE

PG4

Metals

_CL ×4

Waters ×124

* Residue conservation analysis

PDB id:

2vq7

Name:

Oxidoreductase

Title:

Bacterial flavin-containing monooxygenase in complex with nadp: native data

Structure:

Flavin-containing monooxygenase. Chain: a, b, c, d. Engineered: yes. Mutation: yes

Source:

Methylophaga sp. Sk1. Organism_taxid: 230105. Expressed in: escherichia coli. Expression_system_taxid: 511693.

Resolution:

2.60Å R-factor: 0.249 R-free: 0.269

Authors:

A.Alfieri,E.Malito,R.Orru,M.W.Fraaije,A.Mattevi

Key ref:

A.Alfieri et al. (2008). Revealing the moonlighting role of NADP in the structure of a flavin-containing monooxygenase. Proc Natl Acad Sci U S A, 105, 6572-6577. PubMed id: 18443301 DOI: 10.1073/pnas.0800859105

Date:

12-Mar-08 Release date: 22-Apr-08

Protein chains

Q83XK4  (Q83XK4_9GAMM) -  Trimethylamine monooxygenase from Methylophaga aminisulfidivorans

Seq: 456 a.a.

Struc: 443 a.a.*

* PDB and UniProt seqs differ at 2 residue positions (black crosses)

Enzyme reactions

• Enzyme class: E.C.1.14.13.148 - trimethylamine monooxygenase.
• Reaction: trimethylamine + NADPH + O2 = trimethylamine N-oxide + NADP⁺ + H2O

trimethylamine (Bound ligand (Het Group name = NAP) corresponds exactly) + NADPH + O2 = trimethylamine N-oxide + NADP(+) + H2O

• Cofactor: Flavoprotein

Molecule diagrams generated from .mol files obtained from the KEGG ftp site

reference

DOI no: 10.1073/pnas.0800859105

Proc Natl Acad Sci U S A 105:6572-6577 (2008)

PubMed id: 18443301

Revealing the moonlighting role of NADP in the structure of a flavin-containing monooxygenase.

A.Alfieri, E.Malito, R.Orru, M.W.Fraaije, A.Mattevi.

ABSTRACT

Flavin-containing monooxygenases (FMOs) are, after cytochromes P450, the most important monooxygenase system in humans and are involved in xenobiotics metabolism and variability in drug response. The x-ray structure of a soluble prokaryotic FMO from Methylophaga sp. strain SK1 has been solved at 2.6-A resolution and is now the protein of known structure with the highest sequence similarity to human FMOs. The structure possesses a two-domain architecture, with both FAD and NADP(+) well defined by the electron density maps. Biochemical analysis shows that the prokaryotic enzyme shares many functional properties with mammalian FMOs, including substrate specificity and the ability to stabilize the hydroperoxyflavin intermediate that is crucial in substrate oxygenation. On the basis of their location in the structure, the nicotinamide ring and the adjacent ribose of NADP(+) turn out to be an integral part of the catalytic site being actively engaged in the stabilization of the oxygenating intermediate. This feature suggests that NADP(H) has a moonlighting role, in that it adopts two binding modes that allow it to function in both flavin reduction and oxygen reactivity modulation, respectively. We hypothesize that a relative domain rotation is needed to bring NADP(H) to these distinct positions inside the active site. Localization of mutations in human FMO3 that are known to cause trimethylaminuria (fish-odor syndrome) in the elucidated FMO structure provides a structural explanation for their biological effects.

Selected figure(s)

Figure 3.
Overall crystal structure of mFMO. (A) Ribbon diagram of the monomer. FAD-binding domain (residues 8–169 and 281–450) is orange and NADP-binding domain (residues 170–280) is green. FAD is shown as yellow sticks and NADP^+ as blue sticks. The positions of the long interdomain loop (residues 44–80), the hinge connecting the two domains, and the polypeptide stretch corresponding to residues 407–415 are outlined. mFMO residues corresponding to TMAU-causing mutations (17) and polymorphisms in hFMO3 (in parentheses) are in red and blue sticks, respectively. The position of a long insert in hFMO3 (residues 238–299; Fig. 1B) is also indicated. It is expected to occupy a surface-exposed position away from the active site. (B) Ribbon representation of the mFMO dimer. One monomer is shown in the same orientation and color as Fig. 3A; the other one is colored gray, with the NADP-binding domain in darker gray.

Figure 5.
The role of NADP^+ in the stabilization of C4a-hydroperoxyflavin intermediate. (A) Modeling experiment in which the hypothetical structure of C4a-hydroperoxyflavin was superimposed to the flavin in mFMO structure. The color code is the same as in Fig. 4B. Hypothetical hydrogen bonds involving the hydroperoxyflavin atoms are shown as blue dashed lines. The accommodation of the additional oxygen atoms of the C4a-adduct would require a shift of ≈1.5 Å of Asn-78 side chain (whose conformation in the native structure is shown as thin black stick). (B) Comparison of the NADP^+-binding mode in S. pombe (Protein Data Bank ID code 2gv8) and Methylophaga FMOs. The picture was obtained by superimposing the Cα atoms of the two proteins and shows the FAD (yellow) and NADP^+ (blue) molecules of mFMO together with FAD and NADP^+ of the S. pombe enzyme (red). The N5 and C4a atoms of the flavin and C4 and C2 atoms of NADP^+ are labeled.

Figures were selected by an automated process.