Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Saccharolobus solfataricus |
Crystallization (Comment) | Organism |
---|---|
crystals of enzyme/CoA complex are grown by hanging drop vapor diffusion in 20 days at 20°C using a well solution containing 0.1 M MES, pH 6.5, and 12% PEG 20,000. Crystals are cryoprotected using well solution supplemented with 30% glycerol | Saccharolobus solfataricus |
Protein Variants | Comment | Organism |
---|---|---|
D29A | reduction in activity by about 2fold | Saccharolobus solfataricus |
D53N | modest reduction in activity | Saccharolobus solfataricus |
E42Q | modest reduction in activity | Saccharolobus solfataricus |
E43Q | modest reduction in activity | Saccharolobus solfataricus |
E76A | activity is reduced to near background levels | Saccharolobus solfataricus |
E76Q | mutation has no effect on activity | Saccharolobus solfataricus |
H36A | mutant shows activity similar to the wild type enzyme | Saccharolobus solfataricus |
H72A | lowered activity | Saccharolobus solfataricus |
H72A | modest reduction in activity | Saccharolobus solfataricus |
H72A/E76Q | activity is reduced to near background levels | Saccharolobus solfataricus |
M121H | mutation results in about a 2fold increase in protein substrate Km and about a 5fold decrease in overall kcat, despite little change in the Km for acetyl-CoA | Saccharolobus solfataricus |
M121Y | mutation results in about a 2fold increase in protein substrate Km and about a 5fold decrease in overall kcat, despite little change in the Km for acetyl-CoA | Saccharolobus solfataricus |
R33A | reduction in activity to near background levels. Km for Ac-CoA is similar to wild type and elevated by about 2fold for protein substrate, whereas the overall kcat is reduced about 5fold relative to the wild type protein | Saccharolobus solfataricus |
S78A | the activity for both mutants is close to wild type | Saccharolobus solfataricus |
S78C | the activity for both mutants is close to wild type | Saccharolobus solfataricus |
Y38S | modest reduction in activity | Saccharolobus solfataricus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
[DNA-binding protein Alba1]-L-lysine16 | P6A, P8A, and G15A mutants of ALBA have Km values similar to wild type ALBA, thus revealing that these residues do not play a significant role of PAT acetylation of ALBA | Saccharolobus solfataricus | |
0.037 | - |
acetyl-CoA | pH 8.0, 75°C, ALBA 11-mer peptide with the sequence VLIGKKPVMNY, mutant enzyme R33A | Saccharolobus solfataricus | |
0.039 | - |
acetyl-CoA | pH 8.0, 75°C, ALBA 11-mer peptide with the sequence VLIGKKPVMNY, mutant enzyme H72A/E76Q | Saccharolobus solfataricus | |
0.048 | - |
acetyl-CoA | pH 8.0, 75°C, ALBA 11-mer peptide with the sequence VLIGKKPVMNY, mutant enzyme M121H | Saccharolobus solfataricus | |
0.048 | - |
acetyl-CoA | pH 8.0, 75°C, ALBA 11-mer peptide with the sequence VLIGKKPVMNY, wild-type enzyme | Saccharolobus solfataricus | |
0.05 | - |
acetyl-CoA | pH 8.0, 75°C, ALBA 11-mer peptide with the sequence VLIGKKPVMNY, mutant enzyme M121Y | Saccharolobus solfataricus | |
0.059 | - |
acetyl-CoA | pH 8.0, 75°C, ALBA 11-mer peptide with the sequence VLIGKKPVMNY, mutant enzyme E76A | Saccharolobus solfataricus | |
0.11 | - |
[DNA-binding protein Alba1]-L-lysine16 | pH 8.0, 75°C, ALBA 11-mer peptide with the sequence VLIGKKPVMNY, wild-type enzyme | Saccharolobus solfataricus | |
0.58 | - |
VLIGKKPVMNY | pH 8.0, 75°C, ALBA 11-mer peptide with the sequence VLIGKKPVMNY, wild-type enzyme | Saccharolobus solfataricus | |
0.91 | - |
VLIGKKPVMNY | pH 8.0, 75°C, ALBA 11-mer peptide with the sequence VLIGKKPVMNY, mutant enzyme H72A/E76Q | Saccharolobus solfataricus | |
0.94 | - |
VLIGKKPVMNY | pH 8.0, 75°C, ALBA 11-mer peptide with the sequence VLIGKKPVMNY, mutant enzyme E76A | Saccharolobus solfataricus | |
1.2 | - |
VLIGKKPVMNY | pH 8.0, 75°C, ALBA 11-mer peptide with the sequence VLIGKKPVMNY, mutant enzyme M121H | Saccharolobus solfataricus | |
1.2 | - |
VLIGKKPVMNY | pH 8.0, 75°C, ALBA 11-mer peptide with the sequence VLIGKKPVMNY, mutant enzyme R33A | Saccharolobus solfataricus | |
1.3 | - |
VLIGKKPVMNY | pH 8.0, 75°C, ALBA 11-mer peptide with the sequence VLIGKKPVMNY, mutant enzyme M121Y | Saccharolobus solfataricus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
acetyl-CoA + [DNA-binding protein Alba1]-L-lysine16 | Saccharolobus solfataricus | the Sulfolobus solfataricus protein acetyltransferase acetylates ALBA, an abundant nonspecific DNA-binding protein, on Lys16 to reduce its DNA affinity, and the Sir2 deacetylase reverses the modification to cause transcriptional repression. This represents a primitive model for chromatin regulation analogous to histone modification in eukaryotes | CoA + [DNA-binding protein Alba1]-N6-acetyl-L-lysine16 | - |
? | |
acetyl-CoA + [DNA-binding protein Alba1]-L-lysine16 | Saccharolobus solfataricus P2 | the Sulfolobus solfataricus protein acetyltransferase acetylates ALBA, an abundant nonspecific DNA-binding protein, on Lys16 to reduce its DNA affinity, and the Sir2 deacetylase reverses the modification to cause transcriptional repression. This represents a primitive model for chromatin regulation analogous to histone modification in eukaryotes | CoA + [DNA-binding protein Alba1]-N6-acetyl-L-lysine16 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Saccharolobus solfataricus | Q97V23 | - |
- |
Saccharolobus solfataricus P2 | Q97V23 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Saccharolobus solfataricus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
acetyl-CoA + VLIGKKPVMNY | - |
Saccharolobus solfataricus | CoA + VLIG-K(Ac)-KPVMNY | - |
? | |
acetyl-CoA + VLIGKKPVMNY | - |
Saccharolobus solfataricus P2 | CoA + VLIG-K(Ac)-KPVMNY | - |
? | |
acetyl-CoA + [DNA-binding protein Alba1]-L-lysine16 | - |
Saccharolobus solfataricus | CoA + [DNA-binding protein Alba1]-N6-acetyl-L-lysine16 | - |
? | |
acetyl-CoA + [DNA-binding protein Alba1]-L-lysine16 | the Sulfolobus solfataricus protein acetyltransferase acetylates ALBA, an abundant nonspecific DNA-binding protein, on Lys16 to reduce its DNA affinity, and the Sir2 deacetylase reverses the modification to cause transcriptional repression. This represents a primitive model for chromatin regulation analogous to histone modification in eukaryotes | Saccharolobus solfataricus | CoA + [DNA-binding protein Alba1]-N6-acetyl-L-lysine16 | - |
? | |
acetyl-CoA + [DNA-binding protein Alba1]-L-lysine16 | - |
Saccharolobus solfataricus P2 | CoA + [DNA-binding protein Alba1]-N6-acetyl-L-lysine16 | - |
? | |
acetyl-CoA + [DNA-binding protein Alba1]-L-lysine16 | the Sulfolobus solfataricus protein acetyltransferase acetylates ALBA, an abundant nonspecific DNA-binding protein, on Lys16 to reduce its DNA affinity, and the Sir2 deacetylase reverses the modification to cause transcriptional repression. This represents a primitive model for chromatin regulation analogous to histone modification in eukaryotes | Saccharolobus solfataricus P2 | CoA + [DNA-binding protein Alba1]-N6-acetyl-L-lysine16 | - |
? |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
75 | - |
assay at | Saccharolobus solfataricus |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
additional information | - |
the enzyme is nearly completely inactive at room temperature | Saccharolobus solfataricus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0067 | - |
VLIGKKPVMNY | pH 8.0, 75°C, ALBA 11-mer peptide with the sequence VLIGKKPVMNY, mutant enzyme E76A | Saccharolobus solfataricus | |
0.0067 | - |
VLIGKKPVMNY | pH 8.0, 75°C, ALBA 11-mer peptide with the sequence VLIGKKPVMNY, mutant enzyme M121Y | Saccharolobus solfataricus | |
0.0083 | - |
VLIGKKPVMNY | pH 8.0, 75°C, ALBA 11-mer peptide with the sequence VLIGKKPVMNY, mutant enzyme R33A | Saccharolobus solfataricus | |
0.0092 | - |
VLIGKKPVMNY | pH 8.0, 75°C, ALBA 11-mer peptide with the sequence VLIGKKPVMNY, mutant enzyme M121H | Saccharolobus solfataricus | |
0.022 | - |
VLIGKKPVMNY | pH 8.0, 75°C, ALBA 11-mer peptide with the sequence VLIGKKPVMNY, mutant enzyme H72A/E76Q | Saccharolobus solfataricus | |
0.0385 | - |
VLIGKKPVMNY | pH 8.0, 75°C, ALBA 11-mer peptide with the sequence VLIGKKPVMNY, wild-type enzyme | Saccharolobus solfataricus | |
0.0385 | - |
[DNA-binding protein Alba1]-L-lysine16 | pH 8.0, 75°C, ALBA 11-mer peptide with the sequence VLIGKKPVMNY, wild-type enzyme | Saccharolobus solfataricus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Saccharolobus solfataricus |
General Information | Comment | Organism |
---|---|---|
physiological function | the Sulfolobus solfataricus protein acetyltransferase acetylates ALBA, an abundant nonspecific DNA-binding protein, on Lys16 to reduce its DNA affinity, and the Sir2 deacetylase reverses the modification to cause transcriptional repression. This represents a primitive model for chromatin regulation analogous to histone modification in eukaryotes | Saccharolobus solfataricus |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0051 | - |
VLIGKKPVMNY | pH 8.0, 75°C, ALBA 11-mer peptide with the sequence VLIGKKPVMNY, mutant enzyme M121Y | Saccharolobus solfataricus | |
0.0069 | - |
VLIGKKPVMNY | pH 8.0, 75°C, ALBA 11-mer peptide with the sequence VLIGKKPVMNY, mutant enzyme R33A | Saccharolobus solfataricus | |
0.0071 | - |
VLIGKKPVMNY | pH 8.0, 75°C, ALBA 11-mer peptide with the sequence VLIGKKPVMNY, mutant enzyme E76A | Saccharolobus solfataricus | |
0.0077 | - |
VLIGKKPVMNY | pH 8.0, 75°C, ALBA 11-mer peptide with the sequence VLIGKKPVMNY, mutant enzyme M121H | Saccharolobus solfataricus | |
0.024 | - |
VLIGKKPVMNY | pH 8.0, 75°C, ALBA 11-mer peptide with the sequence VLIGKKPVMNY, mutant enzyme H72A/E76Q | Saccharolobus solfataricus | |
0.066 | - |
VLIGKKPVMNY | pH 8.0, 75°C, ALBA 11-mer peptide with the sequence VLIGKKPVMNY, wild-type enzyme | Saccharolobus solfataricus | |
0.35 | - |
[DNA-binding protein Alba1]-L-lysine16 | pH 8.0, 75°C, ALBA 11-mer peptide with the sequence VLIGKKPVMNY, wild-type enzyme | Saccharolobus solfataricus |