Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21 | Pyrococcus furiosus |
Crystallization (Comment) | Organism |
---|---|
mutant enzyme R170A/R220A/Y227F, hanging-drop vapour-diffusion method, the structure is determined at 2.35 A resolution, stable homotetrameric structure, a dimeric form of the enzyme is constructed and its crystal structure is solved at a resolution of 1.70 A | Pyrococcus furiosus |
Protein Variants | Comment | Organism |
---|---|---|
R170A/R220A/Y227F | mutant enzyme occurs as dimer | Pyrococcus furiosus |
R170A/R220A/Y227F/R448E | mutant enzyme occurs as monomer, melting temperature is decreased form 110°C for the wild-type enzyme to 73-75°C for the mutant enzyme | Pyrococcus furiosus |
R170A/R220A/Y227F/R449R | mutant enzyme occurs as monomer, melting temperature is decreased form 110°C for the wild-type enzyme to 73-75°C for the mutant enzyme | Pyrococcus furiosus |
R170A/R220A/Y227F/R459G | mutant enzyme occurs as monomer, melting temperature is decreased form 110°C for the wild-type enzyme to 73-75°C for the mutant enzyme | Pyrococcus furiosus |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
23600 | - |
wild-type enzyme tetramer, gel filtration | Pyrococcus furiosus |
50000 | - |
mutant enzyme R170A/R220A/Y227F/R459G, monomer, gel filtration | Pyrococcus furiosus |
55000 | - |
2 * 55000, SDS-PAGE after heating at 95°C,mutant enzyme R170A/R220A/Y227F | Pyrococcus furiosus |
71000 | - |
mutant enzyme R170A/R220A/Y227F/R448E, monomer, gel filtration | Pyrococcus furiosus |
96000 | - |
mutant enzyme R170A/R220A/Y227F/R449R, monomer, gel filtration | Pyrococcus furiosus |
136000 | - |
mutant enzyme R170A/R220A/Y227F, dimer, gel filtration | Pyrococcus furiosus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pyrococcus furiosus | Q51723 | - |
- |
Purification (Comment) | Organism |
---|---|
wild-type and mutant enzyme R170A/R220A/Y227F | Pyrococcus furiosus |
Subunits | Comment | Organism |
---|---|---|
dimer | 2 * 55000, SDS-PAGE after heating at 95°C,mutant enzyme R170A/R220A/Y227F | Pyrococcus furiosus |
monomer | 1 * 50000, mutant enzyme R170A/R220A/Y227F/R459G, SDS-PAGE | Pyrococcus furiosus |
monomer | 1 * 71000, mutant enzyme R170A/R220A/Y227F/R448E, SDS-PAGE | Pyrococcus furiosus |
monomer | 1 * 96000, mutant enzyme R170A/R220A/Y227F/R449R, SDS-PAGE | Pyrococcus furiosus |
tetramer | wild-type enzyme | Pyrococcus furiosus |
Synonyms | Comment | Organism |
---|---|---|
BGLPf | - |
Pyrococcus furiosus |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
additional information | - |
oligomerization contributes to thermostability | Pyrococcus furiosus |
70 | 80 | mutant enzymes R170A/R220A/Y227F/R448E, R170A/R220A/Y227F/R449R or R170A/R220A/Y227F/R459G are immediately inactivated | Pyrococcus furiosus |
73 | 75 | melting temperature of mutant enzymes R170A/R220A/Y227F/R448E, R170A/R220A/Y227F/R449R or R170A/R220A/Y227F/R459G | Pyrococcus furiosus |
102 | - |
Tm-value of mutant enzyme R170A/R220A/Y227F | Pyrococcus furiosus |
110 | - |
melting temperature of tetrameric wild-type enzyme | Pyrococcus furiosus |