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Literature summary extracted from

  • Sgraja, T.; Smith, T.K.; Hunter, W.N.
    Structure, substrate recognition and reactivity of Leishmania major mevalonate kinase (2007), BMC Struct. Biol., 7, 20.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.1.36 DNA and amino acid sequence determination and anaylsis, overexpression in a procyclic form of Trypanosoma brucei, and expression of His-tagged wild-type enzyme in Escherichia coli strain BL21(DE3), and of His-tagged selenomethionine-labeled enzyme in Escherichia coli strain B834 Leishmania major
2.7.1.36 DNA and amino acid sequence determination and anaylsis, overexprssion in a procyclic form of Trypanosoma brucei Trypanosoma brucei
2.7.1.36 expression in Trypanosoma brucei Leishmania major

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.7.1.36 free enzyme and in complex with mevalonate, 1.75 A and 1.9 A resolution, respectively. The mevalonate binds in a deep cavity lined by highly conserved residues. His25 is key for binding and for discrimination of (R)- over (S)-mevalonate, with the main chain amide interacting with the C3 hydroxyl group of (R)-mevalonate, and the side chain contributing, together with Val202 and Thr283, to the construction of a hydrophobic binding site for the C3 methyl substituent. The C5 hydroxyl, where phosphorylation occurs, points towards catalytic residues, Lys18 and Asp155 Leishmania major
2.7.1.36 purified recombinant wild-type and selenomethionine-labeled enzyme as apoenzyme and in complex with (R)-mevalonate, hanging drop vapor diffusion method, 0.002 ml of 7.5 mg/ml protein in 10 mM Tris-HCl, pH 8.5, 20 mM NaCl, and 1 mM DTT, in presence of 3-25 mM adenosine 5'-(beta,gamma-imino)triphosphate, 6-50 mM (R)-mevalonate, 10 mM Mg2+, mixing with reservoir solution containing 1.15 M sodium citrate, pH 6.2, 18°C, X-ray diffraction structure determination and analysis at 1.75-1.9 A resolution, single-wavelength anomalous dispersion, molecular replacement fails Leishmania major

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.1.36 Mg2+
-
 Leishmania major

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.7.1.36 ATP + (R)-mevalonate Trypanosoma brucei
-
 ADP + (R)-5-phosphomevalonate
-
 r
2.7.1.36 ATP + (R)-mevalonate Leishmania major
-
 ADP + (R)-5-phosphomevalonate
-
 r

Organism

EC Number Organism UniProt Comment Textmining
2.7.1.36 Leishmania major
-
-
-
2.7.1.36 Leishmania major Q4Q6K7
-
-
2.7.1.36 Trypanosoma brucei
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.1.36 recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, dialysis, and anion exchange chromatography to homogeneity Leishmania major

Reaction

EC Number Reaction Comment Organism Reaction ID
2.7.1.36 ATP + (R)-mevalonate = ADP + (R)-5-phosphomevalonate catalytic reaction mechanism, substrate binding structure involves key residue His25 in a deep cavity lined by highly conserved residues, Lys18, Asp155, Val202, and Thr283 are involved in catalysis Leishmania major
a

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.7.1.36 additional information
-
 development of a highly sensitive radioactive assay Leishmania major

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.1.36 ATP + (R)-mevalonate
-
 Trypanosoma brucei ADP + (R)-5-phosphomevalonate
-
 r
2.7.1.36 ATP + (R)-mevalonate
-
 Leishmania major ADP + (R)-5-phosphomevalonate
-
 r

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.7.1.36 30
-
 assay ta Leishmania major

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.1.36 7
-
 assay at Leishmania major

Cofactor

EC Number Cofactor Comment Organism Structure
2.7.1.36 ADP
-
 Trypanosoma brucei
2.7.1.36 ADP
-
 Leishmania major
2.7.1.36 ATP
-
 Trypanosoma brucei
2.7.1.36 ATP helix alpha2 and the preceding polypeptide adopt a conformation impeding access to the nucleotide triphosphate binding site suggesting that a conformational rearrangement is required to allow ATP binding, the ATP binding structure is distinct from related enzymes, overview Leishmania major