Literature summary for 5.1.99.4 extracted from

Bhaumik, P.; Schmitz, W.; Hassinen, A.; Hiltunen, J.K.; Conzelmann, E.; Wierenga, R.K.
The catalysis of the 1,1-proton transfer by alpha-methyl-acyl-CoA racemase is coupled to a movement of the fatty acyl moiety over a hydrophobic, methionine-rich surface (2007), J. Mol. Biol., 367, 1145-1161.

Cloned(Commentary)

Cloned (Comment)	Organism
overexpression in Escherichia coli	Mycobacterium tuberculosis

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant wild-type enzyme in complex with acetyl-CoA, acetoacetyl-CoA, (R,S)-ibuprofenoyl-CoA, (2S)-methylmyristoyl-CoA, or (2R)-methylmyristoyl-CoA, 22°C, hanging-drop vapor-diffusion method, with reservoir solution containing 1.52 M ammonium phosphate, 10 mM BaCl2 at pH 7.0 in 0.0066 ml drops containing a 1:1 v/v mixture of reservoir and protein solution, the latter containing 6 mg/ml in 10 mM potassium phosphate buffer, pH 8.8, X-ray diffraction structure determination and analysis at 1.6-2.3 A resolution	Mycobacterium tuberculosis

Crystallization (Comment)

Organism

purified recombinant wild-type enzyme in complex with acetyl-CoA, acetoacetyl-CoA, (R,S)-ibuprofenoyl-CoA, (2S)-methylmyristoyl-CoA, or (2R)-methylmyristoyl-CoA, 22°C, hanging-drop vapor-diffusion method, with reservoir solution containing 1.52 M ammonium phosphate, 10 mM BaCl2 at pH 7.0 in 0.0066 ml drops containing a 1:1 v/v mixture of reservoir and protein solution, the latter containing 6 mg/ml in 10 mM potassium phosphate buffer, pH 8.8, X-ray diffraction structure determination and analysis at 1.6-2.3 A resolution

Mycobacterium tuberculosis

Inhibitors

Inhibitors	Comment	Organism
(25-R,S)-3,7,12-trihydroxycoprostanoic acid	competitive inhibition	Mycobacterium tuberculosis
(R)-2-methylmyristoyl CoA	binding mode	Mycobacterium tuberculosis
(R,S)-ibuprofenoyl-CoA	competitive inhibition	Mycobacterium tuberculosis
(S)-2-methylmyristoyl CoA	binding mode	Mycobacterium tuberculosis
(S)-ibuprofenoyl-CoA	competitive inhibition	Mycobacterium tuberculosis
acetoacetyl-CoA	-	Mycobacterium tuberculosis
acetyl-CoA	-	Mycobacterium tuberculosis

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
(2S)-2-methylacyl-CoA	Mycobacterium tuberculosis	-	(2R)-2-methylacyl-CoA	-	r

Organism

Organism	UniProt	Comment	Textmining
Mycobacterium tuberculosis	O06543	-	-

Reaction

Reaction	Comment	Organism	Reaction ID
(2S)-2-methylacyl-CoA = (2R)-2-methylacyl-CoA	reaction mechanism, overview, the catalysis of the 1,1-proton transfer by alpha-methyl-acyl-CoA racemase is coupled to a movement of the fatty acyl moiety over a hydrophobic, methionine-rich surface, the mechanism involves the acid/base-pair residues His126 and Asp156, active site geometry, the thioester oxygen atom and the 2-methyl group are in a cisconformation with respect to each other, the thioester oxygen atom fits into an oxyanion hole and the 2-methyl group points into a hydrophobic pocket, Asp127A is a prominent residue in the active site and is located at the bottom of the catalytic site	Mycobacterium tuberculosis	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
(25S)-3,7,12-trihydroxycoprostanoic acid	-	Mycobacterium tuberculosis	(25R)-3,7,12-trihydroxycoprostanoic acid	-	r
(2S)-2-methylacyl-CoA	-	Mycobacterium tuberculosis	(2R)-2-methylacyl-CoA	-	r
(2S)-2-methylacyl-CoA	substrate binding structure, the S-substrate and the R-substrate bind in an S-pocket and an R-pocket, respectively, the enzyme contains a large number of methionine residues in the acyl binding region, located between the S-pocket and the R-pocket, overview	Mycobacterium tuberculosis	(2R)-2-methylacyl-CoA	-	r
(S)-ibuprofenoyl-CoA	binding mode at the active site, binding structure, overview	Mycobacterium tuberculosis	(R)-ibuprofenoyl-CoA	-	r
(S)-Pristanoyl-CoA	-	Mycobacterium tuberculosis	(R)-Pristanoyl-CoA	-	r
additional information	structure of the enzyme in complex with acetyl-CoA, acetoacetyl-CoA, (R,S)-ibuprofenoyl-CoA, (2S)-methylmyristoyl-CoA, or (2R)-methylmyristoyl-CoA, overview	Mycobacterium tuberculosis	?	-	?

Subunits

Subunits	Comment	Organism
More	structure of the enzyme in complex with acetyl-CoA, acetoacetyl-CoA, (R,S)-ibuprofenoyl-CoA, (2S)-methylmyristoyl-CoA, or (2R)-methylmyristoyl-CoA, overview	Mycobacterium tuberculosis

Synonyms

Synonyms	Comment	Organism
alpha-methylacyl coenzyme A racemase	-	Mycobacterium tuberculosis
MCR	-	Mycobacterium tuberculosis

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
37	-	assay at	Mycobacterium tuberculosis

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
8	-	assay at	Mycobacterium tuberculosis