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Literature summary for 3.1.3.2 extracted from

  • 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 (2010), J. Mol. Biol., 404, 639-649.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli as a His-tagged fusion protein Haemophilus influenzae

Crystallization (Commentary)

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

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 [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

Localization Comment Organism GeneOntology No. Textmining
membrane localised to the bacterial outer membrane Haemophilus influenzae 16020
-

Organism

Organism UniProt Comment Textmining
Haemophilus influenzae P26093
-
-

Purification (Commentary)

Purification (Comment) Organism
using Ni-NTA chromatography Haemophilus influenzae

Substrates and Products (Substrate)

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

Synonyms Comment Organism
e (P4) phosphatase
-
Haemophilus influenzae

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
assay at Haemophilus influenzae

Turnover Number [1/s]

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

pH Optimum Minimum pH Optimum Maximum Comment Organism
5.5
-
assay at Haemophilus influenzae