Literature summary for 6.3.2.49 extracted from

Tsuda, T.; Suzuki, T.; Kojima, S.
Crystallization and preliminary X-ray diffraction analysis of Bacillus subtilis YwfE, an L-amino-acid ligase (2012), Acta Crystallogr. Sect. F, 68, 203-206.

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified wild-type and selenomethionine-labeled YwfE in complex with ADP, MgCl2 and dipeptide L-Ala-L-Gln, hanging drop vapour diffusion method, mixing of 0.002 ml of protein solution containing 8 mg/ml protein, 10 mM ADP, 10 mM MgCl2, 100 mM L-Ala-L-Gln with 0.002 ml reservoir solution, equilibration against 0.5 ml of reservoir solution, the deletion-mutant crystals are grown in reservoir solution consisting of 17% w/v PEG 3350, 0.3 M NaCl, 0.1 M MES-NaOH, pH 6.75, 5% v/v ethylene glycol, 1 mM DTT, X-ray diffraction structure determination and analysis at 1.9 A and 2.8 A, respectively	Bacillus subtilis

Crystallization (Comment)

Organism

purified wild-type and selenomethionine-labeled YwfE in complex with ADP, MgCl2 and dipeptide L-Ala-L-Gln, hanging drop vapour diffusion method, mixing of 0.002 ml of protein solution containing 8 mg/ml protein, 10 mM ADP, 10 mM MgCl2, 100 mM L-Ala-L-Gln with 0.002 ml reservoir solution, equilibration against 0.5 ml of reservoir solution, the deletion-mutant crystals are grown in reservoir solution consisting of 17% w/v PEG 3350, 0.3 M NaCl, 0.1 M MES-NaOH, pH 6.75, 5% v/v ethylene glycol, 1 mM DTT, X-ray diffraction structure determination and analysis at 1.9 A and 2.8 A, respectively

Bacillus subtilis

Protein Variants

Protein Variants	Comment	Organism
additional information	generation of a deletion mutant comprising residues Lys4-Tyr468	Bacillus subtilis

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
55000	-	2 * 55000, SDS-PAGE	Bacillus subtilis
105000	-	gel filtration	Bacillus subtilis

Organism

Organism	UniProt	Comment	Textmining
Bacillus subtilis	P39641	-	-
Bacillus subtilis 168	P39641	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant N-terminally GST-tagged wild-type and mutant enzymes (deletion mutant comprising residues Lys4-Tyr468) from Escherichia coli strain ArcticExpress RIL cells (DE3) by glutathione affinity chromatography, tag cleavage by PreScission protease, dialysis and removal of the tag by glutathione affinity chromatography, followed by ultrafiltration and gel filtration, recombinant selenomethionine-labeled enzyme from Escherichia coli strain B834(DE3) purified by the same procedure	Bacillus subtilis

Purification (Comment)

Organism

recombinant N-terminally GST-tagged wild-type and mutant enzymes (deletion mutant comprising residues Lys4-Tyr468) from Escherichia coli strain ArcticExpress RIL cells (DE3) by glutathione affinity chromatography, tag cleavage by PreScission protease, dialysis and removal of the tag by glutathione affinity chromatography, followed by ultrafiltration and gel filtration, recombinant selenomethionine-labeled enzyme from Escherichia coli strain B834(DE3) purified by the same procedure

Bacillus subtilis

Subunits

Subunits	Comment	Organism
dimer	2 * 55000, SDS-PAGE	Bacillus subtilis

Synonyms

Synonyms	Comment	Organism
BacD	-	Bacillus subtilis
YwfE	-	Bacillus subtilis