Literature summary for 4.2.1.114 extracted from

Jeyakanthan, J.; Drevland, R.M.; Gayathri, D.R.; Velmurugan, D.; Shinkai, A.; Kuramitsu, S.; Yokoyama, S.; Graham, D.E.
Substrate specificity determinants of the methanogen homoaconitase enzyme: structure and function of the small subunit (2010), Biochemistry, 49, 2687-2696.

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Cloned(Commentary)

Cloned (Comment)	Organism
small-subunit HACN protein MJ1271 is expressed in Escherichia coli BL21 CodonPlus (DE3)-RIL cells	Methanocaldococcus jannaschii

Crystallization (Commentary)

Crystallization (Comment)	Organism
small-subunit HACN protein (MJ1271), micro batch method, using 50% (w/v) polyethylene glycol 200 and 0.1 M Tris-HCl, pH 4.6, at 20°C	Methanocaldococcus jannaschii

Protein Variants

Protein Variants	Comment	Organism
R26K	the variant forms an relatively efficient isopropylmalate isomerase enzyme	Methanocaldococcus jannaschii
R26V	the variant forms an relatively efficient isopropylmalate isomerase enzyme	Methanocaldococcus jannaschii
R26V/T27Y	the variant resembles the MJ1277 isopropylmalate isomerase small subunit in its flexible loop sequence but demonstrates the broad substrate specificity of the R26V variant	Methanocaldococcus jannaschii
T27A t	the variant has uniformly lower specificity constants for both isopropylmalate isomerase and methanogen homoaconitase substrates	Methanocaldococcus jannaschii

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.022	-	but-1-ene-1,2,4-tricarboxylic acid	wild type enzyme	Methanocaldococcus jannaschii
0.03	-	(1E)-pent-1-ene-1,2,5-tricarboxylic acid	wild type enzyme	Methanocaldococcus jannaschii
0.036	-	(1E)-hex-1-ene-1,2,6-tricarboxylic acid	wild type enzyme	Methanocaldococcus jannaschii
0.135	-	but-1-ene-1,2,4-tricarboxylic acid	mutant enzyme R26K	Methanocaldococcus jannaschii
0.22	-	but-1-ene-1,2,4-tricarboxylic acid	mutant enzyme R26V	Methanocaldococcus jannaschii
0.22	-	but-1-ene-1,2,4-tricarboxylic acid	mutant enzyme T27A	Methanocaldococcus jannaschii
0.269	-	(1E)-pent-1-ene-1,2,5-tricarboxylic acid	mutant enzyme T27A	Methanocaldococcus jannaschii
0.46	-	but-1-ene-1,2,4-tricarboxylic acid	mutant enzyme R26V/T27Y	Methanocaldococcus jannaschii
0.64	-	(1E)-hex-1-ene-1,2,6-tricarboxylic acid	mutant enzyme R26V/T27Y	Methanocaldococcus jannaschii
0.65	-	(1E)-hex-1-ene-1,2,6-tricarboxylic acid	mutant enzyme T27A	Methanocaldococcus jannaschii
0.66	-	(1E)-hex-1-ene-1,2,6-tricarboxylic acid	mutant enzyme R26V	Methanocaldococcus jannaschii
0.87	-	(1E)-pent-1-ene-1,2,5-tricarboxylic acid	mutant enzyme R26V	Methanocaldococcus jannaschii
1.6	-	(1E)-pent-1-ene-1,2,5-tricarboxylic acid	mutant enzyme R26V/T27Y	Methanocaldococcus jannaschii

Organism

Organism	UniProt	Comment	Textmining
Methanocaldococcus jannaschii	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
Toyopearl SuperQ-650 M column chromatography, Resource S column chromatography, and Superdex 200 gel filtration	Methanocaldococcus jannaschii

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
(1E)-hex-1-ene-1,2,6-tricarboxylic acid + H2O	-	Methanocaldococcus jannaschii	(2R)-hexane-1,2,6-tricarboxyclic acid	-	?
(1E)-pent-1-ene-1,2,5-tricarboxylic acid + H2O	-	Methanocaldococcus jannaschii	(2R)-pentane-1,2,5-tricarboxylic acid	-	?
(2R)-butane-1,2,4-tricarboxylic acid	-	Methanocaldococcus jannaschii	but-1-ene-1,2,4-tricarboxylic acid + H2O	-	?
but-1-ene-1,2,4-tricarboxylic acid + H2O	-	Methanocaldococcus jannaschii	(2R)-butane-1,2,4-tricarboxylic acid	-	?
dihomocitrate	-	Methanocaldococcus jannaschii	cis-(homo)2aconitate + H2O	-	?
additional information	HACN has no detectable activity with citraconate or isopropylmalate	Methanocaldococcus jannaschii	?	-	?
trihomocitrate	-	Methanocaldococcus jannaschii	cis-(homo)3aconitate + H2O	-	?

Synonyms

Synonyms	Comment	Organism
HACN	-	Methanocaldococcus jannaschii
MJ1003	large subunit protein of HACN	Methanocaldococcus jannaschii
MJ1271	small subunit protein of HACN	Methanocaldococcus jannaschii

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
0.48	-	but-1-ene-1,2,4-tricarboxylic acid	mutant enzyme R26V	Methanocaldococcus jannaschii
0.66	-	(1E)-pent-1-ene-1,2,5-tricarboxylic acid	wild type enzyme	Methanocaldococcus jannaschii
0.75	-	but-1-ene-1,2,4-tricarboxylic acid	wild type enzyme	Methanocaldococcus jannaschii
1.43	-	but-1-ene-1,2,4-tricarboxylic acid	mutant enzyme R26K	Methanocaldococcus jannaschii
1.7	-	but-1-ene-1,2,4-tricarboxylic acid	mutant enzyme R26V/T27Y	Methanocaldococcus jannaschii
1.9	-	(1E)-hex-1-ene-1,2,6-tricarboxylic acid	mutant enzyme R26V/T27Y	Methanocaldococcus jannaschii
2.2	-	(1E)-pent-1-ene-1,2,5-tricarboxylic acid	mutant enzyme T27A	Methanocaldococcus jannaschii
2.5	-	(1E)-hex-1-ene-1,2,6-tricarboxylic acid	wild type enzyme	Methanocaldococcus jannaschii
2.5	-	but-1-ene-1,2,4-tricarboxylic acid	mutant enzyme T27A	Methanocaldococcus jannaschii
2.8	-	(1E)-hex-1-ene-1,2,6-tricarboxylic acid	mutant enzyme R26V	Methanocaldococcus jannaschii
4.1	-	(1E)-hex-1-ene-1,2,6-tricarboxylic acid	mutant enzyme T27A	Methanocaldococcus jannaschii
5.8	-	(1E)-pent-1-ene-1,2,5-tricarboxylic acid	mutant enzyme R26V	Methanocaldococcus jannaschii
6.6	-	(1E)-pent-1-ene-1,2,5-tricarboxylic acid	mutant enzyme R26V/T27Y	Methanocaldococcus jannaschii

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Release 2023.1 (January 2023)