Literature summary for 4.4.1.20 extracted from

Niegowski, D.; Kleinschmidt, T.; Ahmad, S.; Qureshi, A.A.; Marback, M.; Rinaldo-Matthis, A.; Haeggstroem, J.Z.
Structure and inhibition of mouse leukotriene C4 synthase (2014), PLoS ONE, 9, e96763.

Application

Application	Comment	Organism
analysis	although structural differences near the active site and along the C-terminal alpha-helix V suggest that the mouse and human enzymes may function differently in vivo, the mouse enzyme is a useful tool in pharmacological research and drug development	Mus musculus
drug development	although structural differences near the active site and along the C-terminal alpha-helix V suggest that the mouse and human enzymes may function differently in vivo, the mouse enzyme is a useful tool in pharmacological research and drug development	Mus musculus

Cloned(Commentary)

Cloned (Comment)	Organism
recombinant enzyme expression in Pichia pastoris strain KM71H	Mus musculus

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified enzyme in apoform or in complex with substrate glutathione or product analogue S-hexyl-GSH, mixing of 0.001 ml of 3.5 mg/ml protein in 0.03% w/v DDM w/v, 20 mM Tris pH 8.0, 100 mM NaCl, and 0.5 mM TCEP, with or without 1 mM GSH, with 0.001 ml of reservoir solution containing 1.8-2.2 M NH4SO4, 0.2 M NaCl and 0.1 M Na cacodylate pH 6.1-6.8, for S-hexyl GSH-enzyme crystals, S-hexyl GSH is added to mother liquor and soaking of apo-crystals, X-ray diffraction structure determination and analysis at 2.65-2.7 A resolution	Mus musculus

Crystallization (Comment)

Organism

purified enzyme in apoform or in complex with substrate glutathione or product analogue S-hexyl-GSH, mixing of 0.001 ml of 3.5 mg/ml protein in 0.03% w/v DDM w/v, 20 mM Tris pH 8.0, 100 mM NaCl, and 0.5 mM TCEP, with or without 1 mM GSH, with 0.001 ml of reservoir solution containing 1.8-2.2 M NH4SO4, 0.2 M NaCl and 0.1 M Na cacodylate pH 6.1-6.8, for S-hexyl GSH-enzyme crystals, S-hexyl GSH is added to mother liquor and soaking of apo-crystals, X-ray diffraction structure determination and analysis at 2.65-2.7 A resolution

Mus musculus

Inhibitors

Inhibitors	Comment	Organism	Structure
2-benzoyl-5-(5-[(4-chlorophenyl)(methyl)amino]pyridine-2-carbonyl)benzoic acid	i.e. TK04, competitive mode of binding versus leukotriene A4	Mus musculus

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
additional information	-	additional information	Michaelis-Menten kinetics	Mus musculus
0.036	-	leukotriene A4	pH 7.8, temperature not specified in the publication, recombinant enzyme	Mus musculus
1.2	-	glutathione	pH 7.8, temperature not specified in the publication, recombinant enzyme	Mus musculus

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
endoplasmic reticulum membrane	integral membrane protein in the outer leaflet of the nuclear envelope and in the endoplasmic reticulum	Mus musculus	5789	-

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
18000	-	3 * 18000	Mus musculus

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
leukotriene A4 + glutathione	Mus musculus	-	leukotriene C4	-	?

Organism

Organism	UniProt	Comment	Textmining
Mus musculus	Q60860	-	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
leukotriene A4 + glutathione	-	Mus musculus	leukotriene C4	-	?
leukotriene A4 + glutathione	glutathione-enzyme binding structure, overview	Mus musculus	leukotriene C4	i.e. (5S)-hydroxy-(6R)-S-glutathionyl-7,9-trans-11,14-cis-eicosatetraenoic acid	?
additional information	binding structure of product analogue S-hexyl GSH to the active site of the enzyme, overview	Mus musculus	?	-	?

Subunits

Subunits	Comment	Organism
trimer	3 * 18000	Mus musculus

Synonyms

Synonyms	Comment	Organism
leukotriene C4 synthase	-	Mus musculus
LT C4 synthase	-	Mus musculus
LTC4S	-	Mus musculus

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
81	-	leukotriene A4	pH 7.8, temperature not specified in the publication, recombinant enzyme	Mus musculus
84	-	glutathione	pH 7.8, temperature not specified in the publication, recombinant enzyme	Mus musculus

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.8	-	assay at	Mus musculus

Ki Value [mM]

Ki Value [mM]	Ki Value maximum [mM]	Inhibitor	Comment	Organism	Structure
0.000037	-	2-benzoyl-5-(5-[(4-chlorophenyl)(methyl)amino]pyridine-2-carbonyl)benzoic acid	pH 7.8, temperature not specified in the publication, recombinant enzyme	Mus musculus

IC50 Value

IC50 Value	IC50 Value Maximum	Comment	Organism	Inhibitor	Structure
0.000135	-	pH 7.8, temperature not specified in the publication, recombinant enzyme	Mus musculus	2-benzoyl-5-(5-[(4-chlorophenyl)(methyl)amino]pyridine-2-carbonyl)benzoic acid

General Information

General Information	Comment	Organism
evolution	the enzyme belongs to the superfamily of membrane-associated proteins in eicosanoid and glutathione metabolism, which consists of six integral membrane proteins. Although structural differences near the active site and along the C-terminal alpha-helix V suggest that the mouse and human enzymes may function differently in vivo	Mus musculus
physiological function	the enzyme catalyzes the conjugation reaction between the fatty acid leukotriene A4 and GSH to form the pro-inflammatory leukotriene C4, an important mediator of asthma	Mus musculus

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
68	-	glutathione	pH 7.8, temperature not specified in the publication, recombinant enzyme	Mus musculus
2300	-	leukotriene A4	pH 7.8, temperature not specified in the publication, recombinant enzyme	Mus musculus