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Literature summary for 4.2.1.11 extracted from

  • Imker, H.J.; Fedorov, A.A.; Fedorov, E.V.; Almo, S.C.; Gerlt, J.A.
    Mechanistic diversity in the RuBisCO superfamily: the enolase in the methionine salvage pathway in Geobacillus kaustophilus (2007), Biochemistry, 46, 4077-4089.
    View publication on PubMed
Search for more literature for 4.2.1.11

Activating Compound

Activating Compound Comment Organism Structure
HCO3- Km about 50% increased in presence of sodium bicarbonate, crystallized enzyme structure complexed with HCO3- in its activated form Geobacillus kaustophilus

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21, pET17b expression vector Geobacillus kaustophilus
expressed in Escherichia coli, PCR-amplified genomic DNA from Bacillus subtilis strain 168 ligated into pET15b encoding an N-terminal six-His tag vector Bacillus subtilis

Crystallization (Commentary)

Crystallization (Comment) Organism
hanging drop method, 17 A resolution, X-ray coordinates and structure factors determined, complexes with phosphate, with Mg2+, with Mg2+ and HCO3-, with Mg2+ and the alternate substrate 2,3-diketohexane 1-phosphate identified, activated enolase carboxylated on Lys173, conserved Lys98 in the N-terminal domain determined for C1 proton abstraction Geobacillus kaustophilus

Protein Variants

Protein Variants Comment Organism
K147A requires a 10fold greater concentration of protein for observation of enolization Geobacillus kaustophilus
K173A detectable activity of about 3% of that of wild-type enolase, retains ability to enolize the desthio substrate Geobacillus kaustophilus
K98A unable to catalyze the enolase reaction Geobacillus kaustophilus
additional information site-directed mutagenesis of active site residues, spectrophotometric activity assay performed with elevated concentrations of the mutant enzymes Geobacillus kaustophilus

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ activity depends on Geobacillus kaustophilus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2,3-diketo-5-methylthiopentane 1-phosphate Geobacillus kaustophilus methionine salvage pathway 2-hydroxy-3-keto-5-methylthiopent-1-ene 1-phosphate
-
 ?
2,3-diketo-5-methylthiopentane 1-phosphate Bacillus subtilis methionine salvage pathway 2-hydroxy-3-keto-5-methylthiopent-1-ene 1-phosphate
-
 ?
2,3-diketo-5-methylthiopentane 1-phosphate Bacillus subtilis 168 methionine salvage pathway 2-hydroxy-3-keto-5-methylthiopent-1-ene 1-phosphate
-
 ?

Organism

Organism UniProt Comment Textmining
Bacillus subtilis P37869
-
-
Bacillus subtilis 168 P37869
-
-
Geobacillus kaustophilus Q5KVE7
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant protein, gel filtration Geobacillus kaustophilus
recombinant protein, gel filtration Bacillus subtilis

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
 spectrophotometric assay described, D-ribulose 1-phosphate and 5-methylthio-D-ribulose 1-phosphate in the presence of limiting 5-methylthio-D-ribulose 1-phosphate dehydratase analyzed, enzyme concentrations from 0.1 to 10 microM used Geobacillus kaustophilus
additional information
-
 spectrophotometric assay described, generation of reaction products determined by NMR Bacillus subtilis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2,3-diketo-5-methylthiopentane 1-phosphate methionine salvage pathway Geobacillus kaustophilus 2-hydroxy-3-keto-5-methylthiopent-1-ene 1-phosphate
-
 ?
2,3-diketo-5-methylthiopentane 1-phosphate methionine salvage pathway Bacillus subtilis 2-hydroxy-3-keto-5-methylthiopent-1-ene 1-phosphate
-
 ?
2,3-diketo-5-methylthiopentane 1-phosphate methionine salvage pathway, stereochemical course of the reaction catalyzed by enolase determined, C1 proton abstraction Geobacillus kaustophilus 2-hydroxy-3-keto-5-methylthiopent-1-ene 1-phosphate
-
 ?
2,3-diketo-5-methylthiopentane 1-phosphate methionine salvage pathway, stereochemical course of the reaction catalyzed by enolase determined, C1 proton abstraction Bacillus subtilis 2-hydroxy-3-keto-5-methylthiopent-1-ene 1-phosphate
-
 ?
2,3-diketo-5-methylthiopentane 1-phosphate methionine salvage pathway Bacillus subtilis 168 2-hydroxy-3-keto-5-methylthiopent-1-ene 1-phosphate
-
 ?
2,3-diketo-5-methylthiopentane 1-phosphate methionine salvage pathway, stereochemical course of the reaction catalyzed by enolase determined, C1 proton abstraction Bacillus subtilis 168 2-hydroxy-3-keto-5-methylthiopent-1-ene 1-phosphate
-
 ?
2,3-diketohexane 1-phosphate alternate substrate, C1 proton abstraction Geobacillus kaustophilus ?
-
 ?
2,3-diketohexane 1-phosphate alternate substrate, C1 proton abstraction Bacillus subtilis ?
-
 ?
2,3-diketohexane 1-phosphate alternate substrate, C1 proton abstraction Bacillus subtilis 168 ?
-
 ?

Subunits

Subunits Comment Organism
More composed of a N-terminal alpha,beta domain and a C-terminal domain consisting of eight alpha,alpha barrels, polypeptides packed as tight dimers Geobacillus kaustophilus

Synonyms

Synonyms Comment Organism
enolase
-
 Geobacillus kaustophilus
enolase
-
 Bacillus subtilis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 assay at Geobacillus kaustophilus