Literature summary for 1.1.1.85 extracted from

Lee, S.G.; Nwumeh, R.; Jez, J.M.
Structure and mechanism of isopropylmalate dehydrogenase from Arabidopsis thaliana: insights on leucine and aliphatic glucosinolate biosynthesis (2016), J. Biol. Chem., 291, 13421-13430.

Search for more literature for 1.1.1.85

Cloned(Commentary)

Cloned (Comment)	Organism
expressed in Escherichia coli BL21(DE3) cells	Arabidopsis thaliana

Crystallization (Commentary)

Crystallization (Comment)	Organism
in complex with either isopropylmalate and Mg2+ or NAD+, hanging drop vapor diffusion method, using 1.0 M ammonium phosphate, 0.1 M imidazole, pH 8.0	Arabidopsis thaliana

Protein Variants

Protein Variants	Comment	Organism
D264N	inactive	Arabidopsis thaliana
D288N	inactive	Arabidopsis thaliana
D292N	the mutant shows strongly reduced catalytic efficiency compared to the wild type enzyme	Arabidopsis thaliana
K232M	inactive	Arabidopsis thaliana
L132A	the mutant shows reduced catalytic efficiency compared to the wild type enzyme	Arabidopsis thaliana
L133A	the mutant shows reduced catalytic efficiency compared to the wild type enzyme	Arabidopsis thaliana
N234A	inactive	Arabidopsis thaliana
N234D	inactive	Arabidopsis thaliana
R136A	inactive	Arabidopsis thaliana
R136K	the mutant shows very strongly reduced catalytic efficiency compared to the wild type enzyme	Arabidopsis thaliana
R146A	the mutant shows strongly reduced catalytic efficiency compared to the wild type enzyme	Arabidopsis thaliana
R146K	the mutant shows strongly reduced catalytic efficiency compared to the wild type enzyme	Arabidopsis thaliana
R174A	inactive	Arabidopsis thaliana
R174K	the mutant shows strongly reduced catalytic efficiency compared to the wild type enzyme	Arabidopsis thaliana
V235A	the mutant shows reduced catalytic efficiency compared to the wild type enzyme	Arabidopsis thaliana
Y181A	the mutant shows strongly reduced catalytic efficiency compared to the wild type enzyme	Arabidopsis thaliana
Y181F	the mutant shows reduced catalytic efficiency compared to the wild type enzyme	Arabidopsis thaliana
Y181H	the mutant shows reduced catalytic efficiency compared to the wild type enzyme	Arabidopsis thaliana

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.0035	-	(2R,3S)-3-isopropylmalate	wild type enzyme, at pH 7.5 and 25°C	Arabidopsis thaliana
0.0063	-	(2R,3S)-3-isopropylmalate	mutant enzyme Y181H, at pH 7.5 and 25°C	Arabidopsis thaliana
0.0074	-	(2R,3S)-3-isopropylmalate	mutant enzyme Y181F, at pH 7.5 and 25°C	Arabidopsis thaliana
0.0083	-	(2R,3S)-3-isopropylmalate	mutant enzyme Y181A, at pH 7.5 and 25°C	Arabidopsis thaliana
0.0114	-	(2R,3S)-3-isopropylmalate	mutant enzyme V235A, at pH 7.5 and 25°C	Arabidopsis thaliana
0.0242	-	(2R,3S)-3-isopropylmalate	mutant enzyme L132A, at pH 7.5 and 25°C	Arabidopsis thaliana
0.0278	-	(2R,3S)-3-isopropylmalate	mutant enzyme D292N, at pH 7.5 and 25°C	Arabidopsis thaliana
0.0707	-	(2R,3S)-3-isopropylmalate	mutant enzyme R146A, at pH 7.5 and 25°C	Arabidopsis thaliana
0.119	-	(2R,3S)-3-isopropylmalate	mutant enzyme R146K, at pH 7.5 and 25°C	Arabidopsis thaliana
0.125	-	(2R,3S)-3-isopropylmalate	mutant enzyme L133A, at pH 7.5 and 25°C	Arabidopsis thaliana
0.753	-	(2R,3S)-3-isopropylmalate	mutant enzyme R174K, at pH 7.5 and 25°C	Arabidopsis thaliana
3.73	-	(2R,3S)-3-isopropylmalate	mutant enzyme R136K, at pH 7.5 and 25°C	Arabidopsis thaliana

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
(2R,3S)-3-isopropylmalate + NAD+	Arabidopsis thaliana	-	4-methyl-2-oxovalerate + CO2 + NADH + H+	-	?

Organism

Organism	UniProt	Comment	Textmining
Arabidopsis thaliana	P93882	-	-

Purification (Commentary)

Purification (Comment)	Organism
Ni-NTA column chromatography and Superdex 200 gel filtration	Arabidopsis thaliana

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
(2R,3S)-3-isopropylmalate + NAD+	-	Arabidopsis thaliana	4-methyl-2-oxovalerate + CO2 + NADH + H+	-	?

Synonyms

Synonyms	Comment	Organism
IPMDH2	-	Arabidopsis thaliana
isopropylmalate dehydrogenase	-	Arabidopsis thaliana

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
0.003	-	(2R,3S)-3-isopropylmalate	mutant enzyme D292N, at pH 7.5 and 25°C	Arabidopsis thaliana
0.017	-	(2R,3S)-3-isopropylmalate	mutant enzyme Y181A, at pH 7.5 and 25°C	Arabidopsis thaliana
0.023	-	(2R,3S)-3-isopropylmalate	mutant enzyme R146K, at pH 7.5 and 25°C	Arabidopsis thaliana
0.03	-	(2R,3S)-3-isopropylmalate	mutant enzyme R146A, at pH 7.5 and 25°C	Arabidopsis thaliana
0.033	-	(2R,3S)-3-isopropylmalate	mutant enzyme R136K, at pH 7.5 and 25°C	Arabidopsis thaliana
0.233	-	(2R,3S)-3-isopropylmalate	mutant enzyme Y181F, at pH 7.5 and 25°C	Arabidopsis thaliana
0.47	-	(2R,3S)-3-isopropylmalate	mutant enzyme L132A, at pH 7.5 and 25°C	Arabidopsis thaliana
0.583	-	(2R,3S)-3-isopropylmalate	mutant enzyme Y181H, at pH 7.5 and 25°C	Arabidopsis thaliana
1.62	-	(2R,3S)-3-isopropylmalate	mutant enzyme R174K, at pH 7.5 and 25°C	Arabidopsis thaliana
2	-	(2R,3S)-3-isopropylmalate	mutant enzyme V235A, at pH 7.5 and 25°C	Arabidopsis thaliana
3.1	-	(2R,3S)-3-isopropylmalate	mutant enzyme L133A, at pH 7.5 and 25°C	Arabidopsis thaliana
4.57	-	(2R,3S)-3-isopropylmalate	wild type enzyme, at pH 7.5 and 25°C	Arabidopsis thaliana

Cofactor

Cofactor	Comment	Organism	Structure
NAD+	-	Arabidopsis thaliana

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
0.009	-	(2R,3S)-3-isopropylmalate	mutant enzyme R136K, at pH 7.5 and 25°C	Arabidopsis thaliana
0.109	-	(2R,3S)-3-isopropylmalate	mutant enzyme D292N, at pH 7.5 and 25°C	Arabidopsis thaliana
0.192	-	(2R,3S)-3-isopropylmalate	mutant enzyme R146K, at pH 7.5 and 25°C	Arabidopsis thaliana
0.429	-	(2R,3S)-3-isopropylmalate	mutant enzyme R146A, at pH 7.5 and 25°C	Arabidopsis thaliana
1.92	-	(2R,3S)-3-isopropylmalate	mutant enzyme Y181A, at pH 7.5 and 25°C	Arabidopsis thaliana
2.14	-	(2R,3S)-3-isopropylmalate	mutant enzyme R174K, at pH 7.5 and 25°C	Arabidopsis thaliana
19.29	-	(2R,3S)-3-isopropylmalate	mutant enzyme L132A, at pH 7.5 and 25°C	Arabidopsis thaliana
24.83	-	(2R,3S)-3-isopropylmalate	mutant enzyme L133A, at pH 7.5 and 25°C	Arabidopsis thaliana
32.32	-	(2R,3S)-3-isopropylmalate	mutant enzyme Y181F, at pH 7.5 and 25°C	Arabidopsis thaliana
91.21	-	(2R,3S)-3-isopropylmalate	mutant enzyme Y181H, at pH 7.5 and 25°C	Arabidopsis thaliana
180.3	-	(2R,3S)-3-isopropylmalate	mutant enzyme V235A, at pH 7.5 and 25°C	Arabidopsis thaliana
1303	-	(2R,3S)-3-isopropylmalate	wild type enzyme, at pH 7.5 and 25°C	Arabidopsis thaliana

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Release 2023.1 (January 2023)