Activating Compound | Comment | Organism | Structure |
---|---|---|---|
additional information | is activated through two steps of intramolecular autoproteolysis, the first mediated by a serine residue, and the second by a glutamate, which releases the pro-segment and produces the active enzyme | Pseudomonas sp. GK16 |
Cloned (Comment) | Organism |
---|---|
cloned in Escherichia coli BL21/DE3 using he pET23d plasmids harboring the cloned mature cephalosporin acylase gene from Pseudomonas sp. GK16 and the mutant genes E159M, D161N, D161L, L165N, Q168P, and L379N. The mutant proteins are overexpressed and purified | Pseudomonas sp. GK16 |
Crystallization (Comment) | Organism |
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the crystals of Y202L mutants including intermediates, Y202L-I, Y202-II and Y202L-II, and Q168P and L379N are grown at 22°C using the hanging-drop vapor diffusion method. The three dimensional structures are determined at 2-2.5 A resolution. The pro-segments of the mutants undergo dynamic conformational changes during activation and adopt different loop conformations from one another. The autoproteolytic site is found to form catalytically competent conformation with a solvent water molecule, which is essentially conserved in the cephalosporin acylase mutants | Pseudomonas sp. GK16 |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas sp. GK16 | A4ZVL3 | - |
- |
Purification (Comment) | Organism |
---|---|
overexpressed mutant protein | Pseudomonas sp. GK16 |
Synonyms | Comment | Organism |
---|---|---|
cephalosporin acylase | a member of the N-terminal nucleophile hydrolase family | Pseudomonas sp. GK16 |