Cloned (Comment) | Organism |
---|---|
S226P/R228S/S176A with an N-terminal polyhistidine tag is overexpressed in Escherichia coli | Saccharomonospora viridis |
Crystallization (Comment) | Organism |
---|---|
hanging drop vapor diffusion method at 20°C, crystal structure of the inactive form of a Cut190 mutant, S226P/R228S/S176A, in complex with calcium ions and/or substrates | Saccharomonospora viridis |
Protein Variants | Comment | Organism |
---|---|---|
S226P/R228S | increase in PET degradation, improved activity and thermostability | Saccharomonospora viridis |
S226P/R228S/S176A | inactive mutant enzyme | Saccharomonospora viridis |
General Stability | Organism |
---|---|
stability is regulated by Ca2+ binding. One Ca2+ binds to the enzyme, which induces large conformational changes in several loop regions to stabilize an open conformation | Saccharomonospora viridis |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | three calcium ions bind to S226P/R228S/S176A. Ca2+ binding induces the pocket to open, enabling the substrate to access the pocket more easily. Molecular dynamics simulations suggest that a postreaction state in the engaged form presumably exists between the experimentally observed forms, indicating that the substrate would be cleaved in the engaged form and then requires the enzyme to change to the open form to release the product, a process that Ca2+ can greatly accelerate | Saccharomonospora viridis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Saccharomonospora viridis | W0TJ64 | - |
- |
Saccharomonospora viridis AHK190 | W0TJ64 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Saccharomonospora viridis |
Synonyms | Comment | Organism |
---|---|---|
Cut190 | - |
Saccharomonospora viridis |