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Literature summary extracted from
- Endopeptidase Clp: ATP-dependent Clp protease from Escherichia coli (1994), Methods Enzymol., 244, 314-331.
General Stability
| EC Number | General Stability | Organism |
|---|---|---|
| 3.4.21.92 | ATP and nonhydrolyzable analogs of ATP stabilize subunit ClpA | Escherichia coli |
| 3.4.21.92 | 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 |
| 3.4.21.92 | In buffers containing Triton X-100, subunit ClpA is stable for several days | Escherichia coli |
| 3.4.21.92 | Repeated freezing and thawing leads to loss of activity of subunit ClpA | Escherichia coli |
| 3.4.21.92 | Subunit ClpA is sensitive to freezing | Escherichia coli |
Inhibitors
| EC Number | Inhibitors | Comment | Organism | Structure |
|---|---|---|---|---|
| 3.4.21.92 | diisopropyl fluorophosphate | inhibits both oligopeptidase activity of ClpP and proteinase activity of ClpAP | Escherichia coli |  |
| 3.4.21.92 | fluorosulfonylbenzoyladenosine | - |
Escherichia coli | |
| 3.4.21.92 | High salt concentrations | chloride is much more inhibitory than acetate, divalent anions are also very inhibitory | Escherichia coli | |
| 3.4.21.92 | kappa-casein | strong, competitive | Escherichia coli | |
| 3.4.21.92 | Mg2+ | proteolytic activity of ClpAP is dependent on, but concentrations higher than about 30 mM are inhibitory | Escherichia coli | |
| 3.4.21.92 | Xaa-Tyr-Leu-Tyr-Trp | competitive to succinyl-Leu-Tyr 4-methylcoumarin 7-amide degradation | Escherichia coli | |
Metals/Ions
| EC Number | Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|---|
| 3.4.21.92 | Ca2+ | poor substitute for Mg2+ | Escherichia coli | |
| 3.4.21.92 | Mg2+ | proteolytic activity of ClpAP is dependent on, concentrations higher than about 30 mM are inhibitory | Escherichia coli | |
| 3.4.21.92 | Zn2+ | poor substitute for Mg2+ | Escherichia coli | |
Molecular Weight [Da]
| EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|---|
| 3.4.21.92 | 21000 | - |
1 * 230000, subunit ClpP (12 * 21000, amino acid sequence, subunit of ClpP) | Escherichia coli |
| 3.4.21.92 | 83000 | - |
x * 120000-140000, subunit ClpA, gel filtration, x * 83000, subunit ClpA, amino acid sequence | Escherichia coli |
| 3.4.21.92 | 230000 | - |
1 * 230000, subunit ClpP (12 * 21000, amino acid sequence, subunit of ClpP) | Escherichia coli |
Natural Substrates/ Products (Substrates)
| EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 3.4.21.92 | additional information | Escherichia coli | ClpXP appears to be involved in plasmid maintenance and in phage Mu virulence | ? | - |
? | |
| 3.4.21.92 | 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 | ? | - |
? | |
| 3.4.21.92 | additional information | Escherichia coli CSH100 (ClpA) | ClpXP appears to be involved in plasmid maintenance and in phage Mu virulence | ? | - |
? | |
| 3.4.21.92 | 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
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 3.4.21.92 | Escherichia coli | - |
CSH100 (ClpA) | - |
| 3.4.21.92 | Escherichia coli | - |
wild-type Escherichia coli cells transformed with the multicopy plasmid pWPC9 (ClpP) | - |
| 3.4.21.92 | Escherichia coli CSH100 (ClpA) | - |
CSH100 (ClpA) | - |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 3.4.21.92 | ClpA | Escherichia coli |
| 3.4.21.92 | ClpP | Escherichia coli |
Specific Activity [micromol/min/mg]
| EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|---|
| 3.4.21.92 | additional information | - |
- |
Escherichia coli |
Storage Stability
| EC Number | Storage Stability | Organism |
|---|---|---|
| 3.4.21.92 | -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 |
| 3.4.21.92 | -70°C, ClpP stable | Escherichia coli |
| 3.4.21.92 | -70°C, subunit ClpP stable | Escherichia coli |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 3.4.21.92 | Bacteriophage lambdaO-DNA replication protein + H2O | degraded by ClpXP | Escherichia coli | Hydrolyzed bacteriophage lambdaO-DNA replication protein | - |
? | |
| 3.4.21.92 | beta-Galactosidase fusion proteins + H2O | - |
Escherichia coli | Hydrolyzed beta-galactosidase fusion protein | - |
? | |
| 3.4.21.92 | casein + H2O | alpha-casein | Escherichia coli | small peptides derived from casein | - |
? | |
| 3.4.21.92 | Glucagon + H2O | cleavage at multiple sites | Escherichia coli | Hydrolyzed glucagon | - |
? | |
| 3.4.21.92 | Glucagon + H2O | cleavage at multiple sites | Escherichia coli CSH100 (ClpA) | Hydrolyzed glucagon | - |
? | |
| 3.4.21.92 | 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 | - |
? | |
| 3.4.21.92 | 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 | ? | - |
? | |
| 3.4.21.92 | additional information | when activated by ClpA subunit, ClpP can degrade longer polypeptides and proteins | Escherichia coli | ? | - |
? | |
| 3.4.21.92 | additional information | ClpXP appears to be involved in plasmid maintenance and in phage Mu virulence | Escherichia coli | ? | - |
? | |
| 3.4.21.92 | 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 | ? | - |
? | |
| 3.4.21.92 | 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) | ? | - |
? | |
| 3.4.21.92 | additional information | when activated by ClpA subunit, ClpP can degrade longer polypeptides and proteins | Escherichia coli CSH100 (ClpA) | ? | - |
? | |
| 3.4.21.92 | additional information | ClpXP appears to be involved in plasmid maintenance and in phage Mu virulence | Escherichia coli CSH100 (ClpA) | ? | - |
? | |
| 3.4.21.92 | 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) | ? | - |
? | |
| 3.4.21.92 | Oxidized insulin B-chain + H2O | cleavage at multiple sites | Escherichia coli | Hydrolyzed insulin B-chain | - |
? | |
| 3.4.21.92 | 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 | ? | - |
? | |
| 3.4.21.92 | Succinyl-Ala-Ala-Phe 4-methylcoumarin 7-amide + H2O | ClpP subunit alone | Escherichia coli | Succinyl-Ala-Ala + Phe 4-methylcoumarin 7-amide | - |
? | |
| 3.4.21.92 | Succinyl-Leu-Leu-Val-Tyr 4-methylcoumarin 7-amide + H2O | ClpP subunit alone | Escherichia coli | Succinyl-Leu + Leu + Val-Tyr 4-methylcoumarin 7-amide | - |
? | |
| 3.4.21.92 | Succinyl-Leu-Tyr 4-methylcoumarin 7-amide + H2O | ClpP subunit alone | Escherichia coli | Succinyl-Leu-Tyr + 7-amino-4-methylcoumarin | - |
? | |
Subunits
| EC Number | Subunits | Comment | Organism |
|---|---|---|---|
| 3.4.21.92 | ? | 1 * 230000, subunit ClpP (12 * 21000, amino acid sequence, subunit of ClpP) | Escherichia coli |
| 3.4.21.92 | ? | x * 120000-140000, subunit ClpA, gel filtration, x * 83000, subunit ClpA, amino acid sequence | Escherichia coli |
| 3.4.21.92 | ? | 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 |
| 3.4.21.92 | More | bacteria, tomatoes and Trypanosomes all contain genes for a large protein with extensive homology to the regulartory subunit, ClpA | Escherichia coli |
| 3.4.21.92 | More | enzyme consists of two components: ClpP and ClpA or ClpX | Escherichia coli |
| 3.4.21.92 | More | ClpX participates with ClpP in the rapid and specific degradation of the lambda O protein | Escherichia coli |
| 3.4.21.92 | 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 |
| 3.4.21.92 | More | but the combination of ClpX and ClpP has very little activity against alpha-casein | Escherichia coli |
Temperature Range [°C]
| EC Number | Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 3.4.21.92 | 42 | - |
active in degrading alpha-casein up to 42°C, no proteinase activity at 55°C | Escherichia coli |
Temperature Stability [°C]
| EC Number | Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 3.4.21.92 | 10 | - |
inactivation after a short time | Escherichia coli |
Turnover Number [1/s]
| EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 3.4.21.92 | 0.5 | - |
glucagon | value below | Escherichia coli | |
| 3.4.21.92 | 0.5 | - |
oxidized insulin B-chain | value below | Escherichia coli | |
| 3.4.21.92 | 1.17 | - |
ATP | ATPase activity | Escherichia coli | |
| 3.4.21.92 | 2.17 | - |
ATP | proteolysis | Escherichia coli | |
| 3.4.21.92 | 13.3 | - |
Phe-Ala-Pro-His-Met-Ala-Leu-Val-Pro-Val | - |
Escherichia coli | |
pH Optimum
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 3.4.21.92 | additional information | - |
6.3 (ClpA) | Escherichia coli |
| 3.4.21.92 | 7 | - |
succinyl-Leu-Tyr 4-methylcoumarin 7-amide | Escherichia coli |
| 3.4.21.92 | 7.5 | 9.5 | alpha-casein | Escherichia coli |
Cofactor
| EC Number | Cofactor | Comment | Organism | Structure |
|---|---|---|---|---|
| 3.4.21.92 | 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 | |
| 3.4.21.92 | ATP | only ATP and dATP support alpha-casein degradation | Escherichia coli | |
| 3.4.21.92 | dATP | only ATP and dATP support alpha-casein degradation | Escherichia coli | |
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