Literature summary for 1.1.1.37 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.

Cloned(Commentary)

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

Crystallization (Commentary)

Crystallization (Comment)	Organism
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

Crystallization (Comment)

Organism

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]

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

Natural Substrates/ Products (Substrates)

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

Organism

Organism	UniProt	Comment	Textmining
Thermus thermophilus	P10584	-	-

Purification (Commentary)

Purification (Comment)	Organism
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]

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

Substrates and Products (Substrate)

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

Subunits

Subunits	Comment	Organism
homodimer	2 * 36874.5, recombinant enzyme, sequence calculation	Thermus thermophilus
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

Synonyms	Comment	Organism
MDH	-	Thermus thermophilus

Temperature Optimum [°C]

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

Temperature Range [°C]

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

Temperature Stability [°C]

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

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
10	-	-	Thermus thermophilus

pH Range

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

Cofactor

Cofactor	Comment	Organism	Structure
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

General Information	Comment	Organism
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