Any feedback?
Literature summary for 3.4.21.92 extracted from
- Endopeptidase Clp: ATP-dependent Clp protease from Escherichia coli (1994), Methods Enzymol., 244, 314-331.
General Stability
| General Stability | Organism |
|---|---|
| ATP and nonhydrolyzable analogs of ATP stabilize subunit ClpA | Escherichia coli |
| Dilution, particularly in plastic tubes inactivates, stabilization by including 0.05% v/v Triton X-100 and 0.1 M KCl stabilizes subunit ClpA | Escherichia coli |
| In buffers containing Triton X-100, subunit ClpA is stable for several days | Escherichia coli |
| Repeated freezing and thawing leads to loss of activity of subunit ClpA | Escherichia coli |
| Subunit ClpA is sensitive to freezing | Escherichia coli |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| diisopropyl fluorophosphate | inhibits both oligopeptidase activity of ClpP and proteinase activity of ClpAP | Escherichia coli |  |
| fluorosulfonylbenzoyladenosine | - |
Escherichia coli | |
| High salt concentrations | chloride is much more inhibitory than acetate, divalent anions are also very inhibitory | Escherichia coli | |
| kappa-casein | strong, competitive | Escherichia coli | |
| Mg2+ | proteolytic activity of ClpAP is dependent on, but concentrations higher than about 30 mM are inhibitory | Escherichia coli | |
| Xaa-Tyr-Leu-Tyr-Trp | competitive to succinyl-Leu-Tyr 4-methylcoumarin 7-amide degradation | Escherichia coli | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Ca2+ | poor substitute for Mg2+ | Escherichia coli | |
| Mg2+ | proteolytic activity of ClpAP is dependent on, concentrations higher than about 30 mM are inhibitory | Escherichia coli | |
| Zn2+ | poor substitute for Mg2+ | Escherichia coli | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 21000 | - |
1 * 230000, subunit ClpP (12 * 21000, amino acid sequence, subunit of ClpP) | Escherichia coli |
| 83000 | - |
x * 120000-140000, subunit ClpA, gel filtration, x * 83000, subunit ClpA, amino acid sequence | Escherichia coli |
| 230000 | - |
1 * 230000, subunit ClpP (12 * 21000, amino acid sequence, subunit of ClpP) | Escherichia coli |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| additional information | Escherichia coli | ClpXP appears to be involved in plasmid maintenance and in phage Mu virulence | ? | - |
? | |
| additional information | Escherichia coli | the high degree of similarity among the ClpA-like proteins suggests that Clp-like proteases are likely to be important participants in energy-dependent proteolysis in prokaryotic and eukaryotic cells | ? | - |
? | |
| additional information | Escherichia coli CSH100 (ClpA) | ClpXP appears to be involved in plasmid maintenance and in phage Mu virulence | ? | - |
? | |
| additional information | Escherichia coli CSH100 (ClpA) | the high degree of similarity among the ClpA-like proteins suggests that Clp-like proteases are likely to be important participants in energy-dependent proteolysis in prokaryotic and eukaryotic cells | ? | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Escherichia coli | - |
CSH100 (ClpA) | - |
| Escherichia coli | - |
wild-type Escherichia coli cells transformed with the multicopy plasmid pWPC9 (ClpP) | - |
| Escherichia coli CSH100 (ClpA) | - |
CSH100 (ClpA) | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| ClpA | Escherichia coli |
| ClpP | Escherichia coli |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| additional information | - |
- |
Escherichia coli |
Storage Stability
| Storage Stability | Organism |
|---|---|
| -70°C, 50 mM Tris-HCl, pH 7.5, 2 mM EDTA, 2 mM DTT, 10% v/v glycerol, stable for extended periods | Escherichia coli |
| -70°C, ClpP stable | Escherichia coli |
| -70°C, subunit ClpP stable | Escherichia coli |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| Bacteriophage lambdaO-DNA replication protein + H2O | degraded by ClpXP | Escherichia coli | Hydrolyzed bacteriophage lambdaO-DNA replication protein | - |
? | |
| beta-Galactosidase fusion proteins + H2O | - |
Escherichia coli | Hydrolyzed beta-galactosidase fusion protein | - |
? | |
| casein + H2O | alpha-casein | Escherichia coli | small peptides derived from casein | - |
? | |
| Glucagon + H2O | cleavage at multiple sites | Escherichia coli | Hydrolyzed glucagon | - |
? | |
| Glucagon + H2O | cleavage at multiple sites | Escherichia coli CSH100 (ClpA) | Hydrolyzed glucagon | - |
? | |
| Leu-Tyr-Leu-Tyr-Trp + H2O | cleavage occurs primarily at Leu3-Tyr4, but significant cleavage also at Tyr2-Leu3 and Leu4-Trp5 bond | Escherichia coli | Leu-Tyr-Leu + Tyr-Trp | - |
? | |
| additional information | ClpP subunit has peptidase activity against very short peptides, with fewer than five amino acid residues in the absence of ClpA and nucleotide | Escherichia coli | ? | - |
? | |
| additional information | when activated by ClpA subunit, ClpP can degrade longer polypeptides and proteins | Escherichia coli | ? | - |
? | |
| additional information | ClpXP appears to be involved in plasmid maintenance and in phage Mu virulence | Escherichia coli | ? | - |
? | |
| additional information | the high degree of similarity among the ClpA-like proteins suggests that Clp-like proteases are likely to be important participants in energy-dependent proteolysis in prokaryotic and eukaryotic cells | Escherichia coli | ? | - |
? | |
| additional information | ClpP subunit has peptidase activity against very short peptides, with fewer than five amino acid residues in the absence of ClpA and nucleotide | Escherichia coli CSH100 (ClpA) | ? | - |
? | |
| additional information | when activated by ClpA subunit, ClpP can degrade longer polypeptides and proteins | Escherichia coli CSH100 (ClpA) | ? | - |
? | |
| additional information | ClpXP appears to be involved in plasmid maintenance and in phage Mu virulence | Escherichia coli CSH100 (ClpA) | ? | - |
? | |
| additional information | the high degree of similarity among the ClpA-like proteins suggests that Clp-like proteases are likely to be important participants in energy-dependent proteolysis in prokaryotic and eukaryotic cells | Escherichia coli CSH100 (ClpA) | ? | - |
? | |
| Oxidized insulin B-chain + H2O | cleavage at multiple sites | Escherichia coli | Hydrolyzed insulin B-chain | - |
? | |
| Phe-Ala-Pro-His-Met-Ala-Leu-Val-Pro-Val + H2O | synthetic polypeptide that corresponds to the 10 amino acids surrounding the in vivo processing site in ClpP subunit | Escherichia coli | ? | - |
? | |
| Succinyl-Ala-Ala-Phe 4-methylcoumarin 7-amide + H2O | ClpP subunit alone | Escherichia coli | Succinyl-Ala-Ala + Phe 4-methylcoumarin 7-amide | - |
? | |
| Succinyl-Leu-Leu-Val-Tyr 4-methylcoumarin 7-amide + H2O | ClpP subunit alone | Escherichia coli | Succinyl-Leu + Leu + Val-Tyr 4-methylcoumarin 7-amide | - |
? | |
| Succinyl-Leu-Tyr 4-methylcoumarin 7-amide + H2O | ClpP subunit alone | Escherichia coli | Succinyl-Leu-Tyr + 7-amino-4-methylcoumarin | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| ? | 1 * 230000, subunit ClpP (12 * 21000, amino acid sequence, subunit of ClpP) | Escherichia coli |
| ? | x * 120000-140000, subunit ClpA, gel filtration, x * 83000, subunit ClpA, amino acid sequence | Escherichia coli |
| ? | ClpP is composed of two superimposed rings of six subunits each, ClpA and ClpP form a tight complex in the presence of MgCl2 and ATP, the ClpAP complex is composed of a dodecamer of ClpP and a hexamer of ClpA | Escherichia coli |
| More | bacteria, tomatoes and Trypanosomes all contain genes for a large protein with extensive homology to the regulartory subunit, ClpA | Escherichia coli |
| More | enzyme consists of two components: ClpP and ClpA or ClpX | Escherichia coli |
| More | ClpX participates with ClpP in the rapid and specific degradation of the lambda O protein | Escherichia coli |
| More | ClpP and ClpA interact to form the active protease, this complex degrades a number of proteins, such as alpha-casein into small peptides and can hydrolyze ATP | Escherichia coli |
| More | but the combination of ClpX and ClpP has very little activity against alpha-casein | Escherichia coli |
Temperature Range [°C]
| Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 42 | - |
active in degrading alpha-casein up to 42°C, no proteinase activity at 55°C | Escherichia coli |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 10 | - |
inactivation after a short time | Escherichia coli |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.5 | - |
glucagon | value below | Escherichia coli | |
| 0.5 | - |
oxidized insulin B-chain | value below | Escherichia coli | |
| 1.17 | - |
ATP | ATPase activity | Escherichia coli | |
| 2.17 | - |
ATP | proteolysis | Escherichia coli | |
| 13.3 | - |
Phe-Ala-Pro-His-Met-Ala-Leu-Val-Pro-Val | - |
Escherichia coli | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| additional information | - |
6.3 (ClpA) | Escherichia coli |
| 7 | - |
succinyl-Leu-Tyr 4-methylcoumarin 7-amide | Escherichia coli |
| 7.5 | 9.5 | alpha-casein | Escherichia coli |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| ATP | cleavage of peptides of more than five amino acids residues by ClpP requires activation by ClpA and either ATP or a nonhydrolyzable analog of ATP such as ATPgammaS or 5'-adenylyl imidodiphosphate | Escherichia coli | |
| ATP | only ATP and dATP support alpha-casein degradation | Escherichia coli | |
| dATP | only ATP and dATP support alpha-casein degradation | Escherichia coli | |
Use of this online version of BRENDA is free under the CC BY 4.0 license. See terms of use for full details.
Release 2023.1 (January 2023)
Legal
Curated at
Member of
