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Literature summary extracted from

  • Bacik, J.P.; Tavassoli, M.; Patel, T.R.; McKenna, S.A.; Vocadlo, D.J.; Khajehpour, M.; Mark, B.L.
    Conformational itinerary of Pseudomonas aeruginosa 1,6-anhydro-N-acetylmuramic acid kinase during its catalytic cycle (2014), J. Biol. Chem., 289, 4504-4514.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.1.170 gene anmK, recombinant expression in Escherichia coli strain BL21(DE3) GOLD Pseudomonas aeruginosa

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.7.1.170 purified recombinant enzyme bound to a nonhydrolyzable ATP analogue (AMPPCP) and 1,6-anhydroMurNAc, hanging drop vapor diffusion method, for enzyme AnmK bound to AMPPCP, 6.5 mg/ml protein in 600 mM NaCl, 2 mM AMPPCP, 4 mM MgCl2, 0.5 mM TCEP, and 20 mM Tris, pH 8.0, is mixed with an equal volume of mother liquor containing 26% PEG 4000, 0.2 M MgCl2, 0.1 M Tris, pH 9.0, for enzyme AnmK bound to AMPPCP and the anhMurNAc sugar, AnmK s first co-crystallized with AMPPCP by mixing equal volumes of mother liquor containing 24% PEG 4000, 0.2 M MgCl2, 0.1 M Tris, pH 8.2, with the enzyme at 6.5 mg/ml in 600 mM NaCl, 2 mM AMPPCP, 4 mM MgCl2, 0.5 mM TCEP, 20 mM Tris, pH 8.0, a single crystal of the AnmK-AMPPCP complex is then transferred to a drop of reservoir buffer supplemented with 8 mM anhMurNAc, 2 mM AMPPCP, and 4 mM MgCl2 and soaked for 1 h, at room temperature, X-ray diffraction structure determination and analysis at 1.67-1.84 A resolution Pseudomonas aeruginosa

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.1.170 Mg2+ required, binding structure, overview Pseudomonas aeruginosa

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.7.1.170 ATP + 1,6-anhydro-N-acetyl-beta-muramate + H2O Pseudomonas aeruginosa
-
 ADP + N-acetyl-beta-muramate 6-phosphate + H+
-
 ?
2.7.1.170 additional information Pseudomonas aeruginosa AnmK is an unusual sugar kinase that both cleaves and phosphorylates its 1,6-anhMurNAc substrate ?
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.7.1.170 Pseudomonas aeruginosa Q9I5Q5 gene anmK
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.1.170 recombinant enzyme from Escherichia coli strain BL21(DE3) GOLD by metal affinity chromatography, gel filtration, and ultrafiltration Pseudomonas aeruginosa

Reaction

EC Number Reaction Comment Organism Reaction ID
2.7.1.170 ATP + 1,6-anhydro-N-acetyl-beta-muramate + H2O = ADP + N-acetylmuramate 6-phosphate reaction mechanism, structure-function analysis, overview. A one-step catalytic mechanism is proposed that involves attack of water at the anomeric center leading to cleavage of the 1,6-anhydro ring and phosphoryl transfer. The reaction inverts the anomeric center of MurNAc. Transient formation of an intermediate (MurNAc) in a two-step mechanism may also be possible. It is also possible that the gamma-phosphate leaves the ATP as a transient metaphosphate prior to the anhydro-bond cleavage Pseudomonas aeruginosa
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.1.170 ATP + 1,6-anhydro-N-acetyl-beta-muramate + H2O
-
 Pseudomonas aeruginosa ADP + N-acetyl-beta-muramate 6-phosphate + H+
-
 ?
2.7.1.170 ATP + 1,6-anhydro-N-acetyl-beta-muramate + H2O sugar substrate binding structure, overview Pseudomonas aeruginosa ADP + N-acetyl-beta-muramate 6-phosphate + H+
-
 ?
2.7.1.170 additional information AnmK is an unusual sugar kinase that both cleaves and phosphorylates its 1,6-anhMurNAc substrate Pseudomonas aeruginosa ?
-
 ?

Subunits

EC Number Subunits Comment Organism
2.7.1.170 More open and closed ligand-bound enzyme structure, overview Pseudomonas aeruginosa

Synonyms

EC Number Synonyms Comment Organism
2.7.1.170 1,6-anhydro-N-acetylmuramic acid kinase
-
 Pseudomonas aeruginosa
2.7.1.170 AnmK
-
 Pseudomonas aeruginosa

Cofactor

EC Number Cofactor Comment Organism Structure
2.7.1.170 ATP
-
 Pseudomonas aeruginosa
2.7.1.170 additional information enzyme AnmK adopts an open conformation in solution in the absence of ligand and that it remains in this open state after binding nonhydrolyzable ATP analogue adenosine 5'-(beta,gamma-methylene)triphosphate (AMPPCP) Pseudomonas aeruginosa

General Information

EC Number General Information Comment Organism
2.7.1.170 evolution althoughAnmKadopts a two-domain fold that is structurally similar to proteins of the hexokinase-hsp70-actin superfamily, 1,6-anhydrosugar kinases are mechanistically unique in that they catalyze both the hydrolysis of the 1,6-anhydro ring and the transfer of the gamma-phosphate group from ATP to O6 of sugar substrates Pseudomonas aeruginosa
2.7.1.170 additional information the peptidoglycan recycling enzyme 1,6-anhydro-N-acetylmuramic acid kinase from Pseudomonas aeruginosa undergoes large conformational changes during its catalytic cycle, with its two domains rotating apart by up to 32° around two hinge regions to expose an active site cleft into which the substrates 1,6-anhydroMurNAc and ATP can bind. Ligand binding at the active site of AnmK coordinates its conformational itinerary Pseudomonas aeruginosa