Literature summary for 6.4.1.2 extracted from

Shen, Y.; Volrath, S.L.; Weatherly, S.C.; Elich, T.D.; Tong, L.
A mechanism for the potent inhibition of eukaryotic acetyl-coenzyme A carboxylase by soraphen A, a macrocyclic polyketide natural product (2004), Mol. Cell, 16, 881-891.

Cloned(Commentary)

Cloned (Comment)	Organism
overexpression of the His-tagged biotin carboxylase domain in Escherichia coli strain BL21(DE3) and as selenomethionyl enzyme in strain B834(DE3)	Saccharomyces cerevisiae

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified selenomethionyl biotin carboxylase domain free and with bound inhibitor soraphen A, sitting drop vapour diffusion method for crystallization of free enzyme: 4°C, the reservoir solution contains 0.1 M Bis-Tris propane, pH 6.0, 23% w/v PEG 3350, 0.2 M NaCl, 0.4 M MgCl2, and 5% glycerol, 12-18 days, followed by microseeding, which is essential, sitting drop vapour diffusion method for crystallization of inhibitor-bound enzyme: 50 mg/ml protein are incubated with 0.88 mM soraphen A at 4°C for 1 h prior to crystallization, the reservoir solution for crystallization at 22°C contains 0.1 M Bis-Tris, pH 5.8, 26% w/v PEG 3350, 0.1 M NaCl, 0.2 M MgCl2, 8% glycerol, and 2 mM DTT, as for the free enzyme microseeding is essential, X-ray diffraction structure determination and analysis at 1.8-2.9 A resolution, selenomethionyl multiwavelength anomalous diffraction method	Saccharomyces cerevisiae

Crystallization (Comment)

Organism

purified selenomethionyl biotin carboxylase domain free and with bound inhibitor soraphen A, sitting drop vapour diffusion method for crystallization of free enzyme: 4°C, the reservoir solution contains 0.1 M Bis-Tris propane, pH 6.0, 23% w/v PEG 3350, 0.2 M NaCl, 0.4 M MgCl2, and 5% glycerol, 12-18 days, followed by microseeding, which is essential, sitting drop vapour diffusion method for crystallization of inhibitor-bound enzyme: 50 mg/ml protein are incubated with 0.88 mM soraphen A at 4°C for 1 h prior to crystallization, the reservoir solution for crystallization at 22°C contains 0.1 M Bis-Tris, pH 5.8, 26% w/v PEG 3350, 0.1 M NaCl, 0.2 M MgCl2, 8% glycerol, and 2 mM DTT, as for the free enzyme microseeding is essential, X-ray diffraction structure determination and analysis at 1.8-2.9 A resolution, selenomethionyl multiwavelength anomalous diffraction method

Saccharomyces cerevisiae

Protein Variants

Protein Variants	Comment	Organism
K73R	the mutant shows resistance to soraphen A	Saccharomyces cerevisiae

Inhibitors

Inhibitors	Comment	Organism	Structure
soraphen A	natural macrocyclic polyketid, inhibition mechanism, the inhibitor binds to an allosteric site in vicinity to the active site at the biotin carboxylase domain dimer interface causing only small coformational changes, binding structure and specificity	Saccharomyces cerevisiae

Organism

Organism	UniProt	Comment	Textmining
Saccharomyces cerevisiae	Q00955	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged biotin carboxylase domain from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, anion exchange chromatography, and gel filtration	Saccharomyces cerevisiae

Synonyms

Synonyms	Comment	Organism
ACC	-	Saccharomyces cerevisiae

Cofactor

Cofactor	Comment	Organism	Structure
ATP	-	Saccharomyces cerevisiae
biotin	-	Saccharomyces cerevisiae