Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli as a His-tagged fusion protein | Haemophilus influenzae |
Crystallization (Comment) | Organism |
---|---|
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. 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 | Haemophilus influenzae |
Protein Variants | Comment | Organism |
---|---|---|
D66N | D66N, a mutant of P4 in which the residue that protonates the leaving group (Asp66) is changed to Asn, is used for structure determination of enzyme-substrate complexes. Structures of D66N complexed with nicotinamide mononucleotide and 5'-AMP are determined at high-resolution limits of 1.35 A and 1.55 A, respectively | Haemophilus influenzae |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.23 | - |
5'-AMP | pH 5.5, 25°C | Haemophilus influenzae | |
0.7 | - |
nicotinamide mononucleotide | pH 5.5, 25°C | Haemophilus influenzae | |
0.8 | - |
3'-AMP | pH 5.5, 25°C | Haemophilus influenzae | |
6 | - |
2'-AMP | pH 5.5, 25°C | Haemophilus influenzae |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
membrane | localised to the bacterial outer membrane | Haemophilus influenzae | 16020 | - |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Haemophilus influenzae | P26093 | - |
- |
Purification (Comment) | Organism |
---|---|
using Ni-NTA chromatography | Haemophilus influenzae |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2'-AMP + H2O | - |
Haemophilus influenzae | adenosine + phosphate | - |
? | |
3'-AMP + H2O | - |
Haemophilus influenzae | adenosine + phosphate | - |
? | |
5'-AMP + H2O | - |
Haemophilus influenzae | adenosine + phosphate | - |
? | |
nicotinamide mononucleotide + H2O | - |
Haemophilus influenzae | nicotinamide riboside + phosphate | - |
? |
Synonyms | Comment | Organism |
---|---|---|
e (P4) phosphatase | - |
Haemophilus influenzae |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Haemophilus influenzae |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.07 | - |
5'-AMP | pH 5.5, 25°C | Haemophilus influenzae | |
0.1 | - |
3'-AMP | pH 5.5, 25°C | Haemophilus influenzae | |
0.11 | - |
2'-AMP | pH 5.5, 25°C | Haemophilus influenzae | |
0.52 | - |
nicotinamide mononucleotide | pH 5.5, 25°C | Haemophilus influenzae |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
5.5 | - |
assay at | Haemophilus influenzae |