Cloned (Comment) | Organism |
---|---|
overexpression of P450cam mutant C334A in Escherichia coli strain BL21(DE3) | Pseudomonas putida |
Crystallization (Comment) | Organism |
---|---|
camphor-free or camphor-bound P450cam mutant C334A in the absence of substrate and at high and low K+ concentration, protein in 50 mM Tris, pH 7.4, with or without 2-4 mM camphor, mixed with crystallization solution containing 50 mM Tris, pH 7.4, and 12-22% PEG 8000 with and without 200 mM K+, sitting drop vapour diffusion method, 6°C, X-ray diffraction structure determination and analysis at 1.50-1.79 A resolution | Pseudomonas putida |
Protein Variants | Comment | Organism |
---|---|---|
C334A | the mutation of P450cam increases the protein stability compared to the wild-type enzyme | Pseudomonas putida |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas putida | P00183 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant P450cam mutant C334A from Escherichia coli strain BL21(DE3) by anion exchange chromatography and gel filtration | Pseudomonas putida |
Subunits | Comment | Organism |
---|---|---|
More | the camphor-free structure is in an open conformation characterized by a water-filled channel created by the retraction of the F and G helices, disorder of the B' helix, and loss of the K+ binding. site. Presence of K+ alone does not alter the open conformation, while camphor alone is sufficient for closure of the channel | Pseudomonas putida |
Synonyms | Comment | Organism |
---|---|---|
P450cam | - |
Pseudomonas putida |