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Literature summary extracted from

  • Knill, T.; Reichelt, M.; Paetz, C.; Gershenzon, J.; Binder, S.
    Arabidopsis thaliana encodes a bacterial-type heterodimeric isopropylmalate isomerase involved in both Leu biosynthesis and the Met chain elongation pathway of glucosinolate formation (2009), Plant Mol. Biol., 71, 227-239.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.2.1.33 full length reading frame of IPMI small subunit 2 fused in frame to the 5' end of the gene encoding the green fluorescent protein in the vector psmGFP4. Construct transiently expressed in tobacco protoplasts and stably transformed into Arabidopsis plants Arabidopsis thaliana
4.2.1.33 recombinant expression of GFP-tagged large and small subunit 1 in Arabidopsis thaliana leaves Arabidopsis thaliana

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
4.2.1.33 chloroplast
-
 Arabidopsis thaliana 9507
-
4.2.1.33 chloroplast presence of different heterodimeric IPMIs in chloroplasts with distinct substrate specificities for Leu or glucosinolate metabolism Arabidopsis thaliana 9507
-
4.2.1.33 chloroplast presence of different heterodimeric IPMIs in chloroplasts with distinct substrate specificities for Leu or glucosinolate metabolism. Import of the IPMI small subunit 2:GFP fusion protein into chloroplasts Arabidopsis thaliana 9507
-
4.2.1.170 chloroplast
-
 Arabidopsis thaliana 9507
-

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
4.2.1.33 (2S)-2-isopropylmalate Arabidopsis thaliana
-
 (2R,3S)-3-isopropylmalate
-
 ?
4.2.1.33 (2S)-2-isopropylmalate Arabidopsis thaliana Col-0
-
 (2R,3S)-3-isopropylmalate
-
 ?
4.2.1.170 2-(2-methylthio)ethylmalate Arabidopsis thaliana
-
 2-(2-methylthio)ethylmaleate + H2O
-
 ?
4.2.1.170 2-(2-methylthio)ethylmalate Arabidopsis thaliana Col-0
-
 2-(2-methylthio)ethylmaleate + H2O
-
 ?
4.2.1.170 2-(3-methylthio)propylmalate Arabidopsis thaliana
-
 3-(3-methylthio)propylmaleate + H2O
-
 ?
4.2.1.170 2-(3-methylthio)propylmalate Arabidopsis thaliana Col-0
-
 3-(3-methylthio)propylmaleate + H2O
-
 ?
4.2.1.170 additional information Arabidopsis thaliana the enzyme catalyzes a two-step reaction via a dehydration of 2-(2-methylthio)ethylmalate and a hydration resulting in an isomerization at position 2/3 to form 3-(2-methylthio)ethylmalate ?
-
 ?
4.2.1.170 additional information Arabidopsis thaliana Col-0 the enzyme catalyzes a two-step reaction via a dehydration of 2-(2-methylthio)ethylmalate and a hydration resulting in an isomerization at position 2/3 to form 3-(2-methylthio)ethylmalate ?
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
4.2.1.33 Arabidopsis thaliana Q94AR8 large subunit
-
4.2.1.33 Arabidopsis thaliana Q94AR8 AND Q9ZW84 large subunit and small subunit 1
-
4.2.1.33 Arabidopsis thaliana Q9LYT7 small subunit
-
4.2.1.33 Arabidopsis thaliana Q9ZW84 small subunit
-
4.2.1.33 Arabidopsis thaliana Q9ZW85 small subunit
-
4.2.1.33 Arabidopsis thaliana Col-0 Q94AR8 AND Q9ZW84 large subunit and small subunit 1
-
4.2.1.170 Arabidopsis thaliana Q94AR8 AND Q9LYT7 large subunit and small subunit 2
-
4.2.1.170 Arabidopsis thaliana Q94AR8 AND Q9ZW85 large subunit and small subunit 3
-
4.2.1.170 Arabidopsis thaliana Col-0 Q94AR8 AND Q9LYT7 large subunit and small subunit 2
-
4.2.1.170 Arabidopsis thaliana Col-0 Q94AR8 AND Q9ZW85 large subunit and small subunit 3
-

Source Tissue

EC Number Source Tissue Comment Organism Textmining
4.2.1.33 leaf
-
 Arabidopsis thaliana
-
4.2.1.33 rosette leaf
-
 Arabidopsis thaliana
-
4.2.1.33 seed
-
 Arabidopsis thaliana
-
4.2.1.33 seedling
-
 Arabidopsis thaliana
-
4.2.1.170 leaf
-
 Arabidopsis thaliana
-
4.2.1.170 seed
-
 Arabidopsis thaliana
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.2.1.33 (2S)-2-isopropylmalate
-
 Arabidopsis thaliana (2R,3S)-3-isopropylmalate
-
 ?
4.2.1.33 (2S)-2-isopropylmalate isomerization via a dehydration and a hydration step Arabidopsis thaliana (2R,3S)-3-isopropylmalate
-
 ?
4.2.1.33 (2S)-2-isopropylmalate
-
 Arabidopsis thaliana Col-0 (2R,3S)-3-isopropylmalate
-
 ?
4.2.1.33 (2S)-2-isopropylmalate isomerization via a dehydration and a hydration step Arabidopsis thaliana Col-0 (2R,3S)-3-isopropylmalate
-
 ?
4.2.1.33 additional information NMR and mass spectrometric compound determination and analysis, overview Arabidopsis thaliana ?
-
 ?
4.2.1.33 additional information NMR and mass spectrometric compound determination and analysis, overview Arabidopsis thaliana Col-0 ?
-
 ?
4.2.1.170 2-(2-methylthio)ethylmalate
-
 Arabidopsis thaliana 2-(2-methylthio)ethylmaleate + H2O
-
 ?
4.2.1.170 2-(2-methylthio)ethylmalate
-
 Arabidopsis thaliana Col-0 2-(2-methylthio)ethylmaleate + H2O
-
 ?
4.2.1.170 2-(3-methylthio)propylmalate
-
 Arabidopsis thaliana 3-(3-methylthio)propylmaleate + H2O
-
 ?
4.2.1.170 2-(3-methylthio)propylmalate
-
 Arabidopsis thaliana Col-0 3-(3-methylthio)propylmaleate + H2O
-
 ?
4.2.1.170 additional information the enzyme catalyzes a two-step reaction via a dehydration of 2-(2-methylthio)ethylmalate and a hydration resulting in an isomerization at position 2/3 to form 3-(2-methylthio)ethylmalate Arabidopsis thaliana ?
-
 ?
4.2.1.170 additional information NMR and mass spectrometric compound determination and analysis, overview Arabidopsis thaliana ?
-
 ?
4.2.1.170 additional information the enzyme catalyzes a two-step reaction via a dehydration of 2-(2-methylthio)ethylmalate and a hydration resulting in an isomerization at position 2/3 to form 3-(2-methylthio)ethylmalate Arabidopsis thaliana Col-0 ?
-
 ?
4.2.1.170 additional information NMR and mass spectrometric compound determination and analysis, overview Arabidopsis thaliana Col-0 ?
-
 ?

Subunits

EC Number Subunits Comment Organism
4.2.1.33 heterodimer
-
 Arabidopsis thaliana

Synonyms

EC Number Synonyms Comment Organism
4.2.1.33 IPMI
-
 Arabidopsis thaliana
4.2.1.33 isopropylmalate dehydratase
-
 Arabidopsis thaliana
4.2.1.33 isopropylmalate isomerase
-
 Arabidopsis thaliana
4.2.1.170 IPMI
-
 Arabidopsis thaliana
4.2.1.170 isopropylmalate dehydratase
-
 Arabidopsis thaliana
4.2.1.170 isopropylmalate isomerase
-
 Arabidopsis thaliana

Expression

EC Number Organism Comment Expression
4.2.1.33 Arabidopsis thaliana in ipmi ssu2-1 plants, absence of mature mRNAs in homozygous plants and the knockout of these genes by T-DNAs down
4.2.1.33 Arabidopsis thaliana in ipmi ssu3-1 plants, absence of mature mRNAs in homozygous plants and the knockout of these genes by T-DNAs down
4.2.1.33 Arabidopsis thaliana in the ipmilsu1-2mutant mRNA levels are reduced to 65 and 50% of the wild-type standard in seedlings and rosette leaves, respectively, while IPMI large subunit 1 transcript levels are reduced to 40 and 27% of wild-type in these tissues of ipmi lsu1-1 plants. Strongest reduction in ipmi lsu1-3 seedlings (21% of wild-type), while the IPMI large subunit 1 mRNA level in ipmi lsu1-3 reaches about 29% of the wild-type level in leaves down

General Information

EC Number General Information Comment Organism
4.2.1.33 malfunction for IPMI small subunit 1, no plants homozygous for the T-DNA allele can be obtained in either analyzed line (ipmi ssu1-1 or ipmi ssu1-2) Arabidopsis thaliana
4.2.1.33 malfunction homozygous ipmi 3-1mutants can be established. In leaves and seeds of the ipmi ssu3-1 mutant few significant changes in amino acid content. Met levels are unchanged in ipmi ssu3-1 and S-methylmethionine can not be detected Arabidopsis thaliana
4.2.1.33 malfunction homozygous ipmi ssu2-1 mutants can be established. Free amino acid content of the leaves and seeds of ipmi ssu2-1 plants is not substantially different from that of the wild-type. S-methylmethionine is not detectable in the ipmi ssu2-1 mutant. Profile of the methionine-derived glucosinolates in leaves of ipmi ssu2-1 does not show substantial variations from that of wild-type plants Arabidopsis thaliana
4.2.1.33 malfunction in leaves, ipmi lsu1-2 plants exhibit the weakest chemical phenotype in comparison to wild-type: transport or storage form of Met is barely detectable. Many more changes between the ipmi lsu1-1 and 1-3 plants and wild-type: accumulation of S-methylmethionine in rosette leaves of both mutants which is accompanied by 2fold increased levels of Met. Average 2-isopropylmalate content of 0.42 and 0.02 mg/g dry weight in leaves of ipmi lsu1-3 and ipmi lsu1-1 mutants, respectively, while this metabolite is undetectable in the ipmi lsu1-2 plants. Relative levels of 2-(3'-methylsulfinyl)propylmalate are 10/1/14 in ipmi lsu1-1, 1-2 and 1-3 plants Arabidopsis thaliana
4.2.1.33 malfunction isopropylmalate isomerase (IPMI) large subunit mutants reveal accumulation of intermediates of both Leu biosynthesis and Met chain elongation, and an altered composition of aliphatic glucosinolates demonstrating the function of this gene in both pathways In contrast, the small subunits appear to be specialized to either Leu biosynthesis, EC 4.2.1.133, or Met chain elongation, EC 4.2.1.170, metabolic profiling, overview. In large subunit single gene knockout mutant IPMI, the remaining IPMI LSU1 transcript is sufficient for some biosynthesis of Met-derived aliphatic glucosinolates and maintaining Leu levels comparable to wild-type plants Arabidopsis thaliana
4.2.1.33 metabolism the enzyme is involved in the leucine biosynthesis. The formation of leucine requires a three-step chain elongation, including a condensation of a 2-oxo acid with acetyl-CoA, an isomerization and an oxidation-decarboxylation, catalyzed by isopropylmalate synthase (IPMS), isopropylmalate isomerase (IPMI, also called a dehydratase) and isopropylmalate dehydrogenase (IPMDH), respectively. Leu biosynthesis is then completed by a transamination step catalyzed by a branched-chain aminotransferase that could be also involved in the formation of the other branched-chain amino acids. The large IPMI subunit is involved in both Leu and glucosinolate metabolism, while the small subunits appear to be specific for each pathway Arabidopsis thaliana
4.2.1.33 physiological function an intact IPMI small subunit 1 gene seems to be crucial for the development of mature viable seeds Arabidopsis thaliana
4.2.1.33 physiological function function of IPMI large subunit 1 in both the biosynthesis of Leu and in the Met chain elongation pathway of glucosinolate formation Arabidopsis thaliana
4.2.1.170 malfunction isopropylmalate isomerase (IPMI) large subunit mutants reveal accumulation of intermediates of both Leu biosynthesis and Met chain elongation, and an altered composition of aliphatic glucosinolates demonstrating the function of this gene in both pathways In contrast, the small subunits appear to be specialized to either Leu biosynthesis, EC 4.2.1.133, or Met chain elongation, EC 4.2.1.170, metabolic profiling, overview Arabidopsis thaliana
4.2.1.170 metabolism the enzyme is involved in methine elongation and in glucosinolate core biosynthesis and side-chain modification. IPMI small subunits are imported into the chloroplast, the localization of the Met chain elongation pathway in these organelles. The large IPMI subunit is involved in both Leu and glucosinolate metabolism, while the small subunits appear to be specific for each pathway. The small subunits IPMI SSU2 and IPMI SSU3 function in Met chain elongation Arabidopsis thaliana