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Literature summary extracted from

  • Hung, C.H.; Hwang, T.S.; Chang, Y.Y.; Luo, H.R.; Wu, S.P.; Hsu, C.H.
    Crystal structures and molecular dynamics simulations of thermophilic malate dehydrogenase reveal critical loop motion for co-substrate binding (2013), PLoS ONE, 8, e83091.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.37 gene mdh, recombinant expression of C-terminally His-tagged enzyme in Escherichia coli strain BL21(DE3) Thermus thermophilus

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.1.1.37 purified recombinant enzyme in apoform and in complex with NAD+, sitting drop vapor diffusion method, mixing 500 nl of 10 mg/ml protein with an equal volume of mother liquor and equilibration against 0.1 ml of the crystallization solution containing 100 mM Tris-HCl, pH 8.2, 22.5% w/v PEG4000, and 200 mM magnesium chloride, at 10°C, 5 days. NAD-bound form crystals are obtained by soaking the apo-form crystal with a 001 ml crystallization reservoir containing 2 mM NAD+ molecule for about 30 min immediately prior X-ray diffraction, X-ray diffraction structure determination and analysis at 1.80-2.36 A resolution, molecular replacement and modelling, overview Thermus thermophilus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.37 61000
-
 recombinant enzyme, gel filtration Thermus thermophilus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.1.37 (S)-malate + NAD+ Thermus thermophilus
-
 oxaloacetate + NADH + H+
-
 r
1.1.1.37 oxaloacetate + NADH + H+ Thermus thermophilus
-
 (S)-malate + NAD+
-
 r

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.37 Thermus thermophilus P10584
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.37 recombinant C-terminally His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration Thermus thermophilus

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.1.1.37 2600
-
 pH 10.0, 37°C Thermus thermophilus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.37 (S)-malate + NAD+
-
 Thermus thermophilus oxaloacetate + NADH + H+
-
 r
1.1.1.37 additional information crystal structures and molecular dynamics simulations of thermophilic malate dehydrogenase reveal critical loop motion for co-substrate binding Thermus thermophilus ?
-
 ?
1.1.1.37 oxaloacetate + NADH + H+
-
 Thermus thermophilus (S)-malate + NAD+
-
 r

Subunits

EC Number Subunits Comment Organism
1.1.1.37 homodimer 2 * 36874.5, recombinant enzyme, sequence calculation Thermus thermophilus
1.1.1.37 More structure analysis, overview. Similar to other MDHs, a protomer of TtMDH has the N-terminal NAD binding domain and C-terminal catalytic domain and consists of 12 helices and 11 beta-strands. The N-terminal NAD binding domain (residue 1-156) is an open twisted structure with the classical Rossmann fold composed of a parallel six-stranded beta-sheet (beta1-beta6) and four alpha-helices (alpha1-alpha4). The C-terminal catalytic domain comprises an antiparallel twisted five-stranded sheet (beta7-beta11) surrounded by eight alpha-helices Thermus thermophilus

Synonyms

EC Number Synonyms Comment Organism
1.1.1.37 MDH
-
 Thermus thermophilus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.37 90
-
-
 Thermus thermophilus

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
1.1.1.37 65 100 high activity within this range Thermus thermophilus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.1.1.37 30 100 high thermal stability of TtMDH, that does not reach a completely denatured state even at near 100°C Thermus thermophilus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.37 10
-
-
 Thermus thermophilus

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.1.1.37 8.5 11 over 50% of maximal activity within this range Thermus thermophilus

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.37 NAD+ a slight movement of the enzymes binding loop and few structural elements around the co-substrate binding packet occurs in the presence of NAD+. The overall structures do not change much and retain all related positions Thermus thermophilus

General Information

EC Number General Information Comment Organism
1.1.1.37 additional information molecular dynamics simulation at higher temperatures were used to reconstruct structures from the crystal structure of TtMDH. At the simulated structure of 80°C, a large change occurs around the active site such that with increasing temperature, a mobile loop is closed to co-substrate binding region. The thermal-induced conformational change of the co-substrate binding loop of TtMDH may contribute to the essential movement of the enzyme for admitting NAD and may benefit the enzyme's activity Thermus thermophilus