Literature summary for 3.4.22.52 extracted from
Vilei, E.M.; Calderara, S.; Anagli, J
Berardi, S.; Hitomi, K.; Maki, M: Carafoli, E.: Functional properties of recombinant calpain I and of mutants lacking domains III and IV of the catalytic subunits (1997), J. Biol. Chem., 727, 25802-25808.
No PubMed abstract available
Cloned(Commentary)
Cloned (Comment) |
Organism |
large catalytic subunit and two of its mutants are expressed in Escherichia coli using the baculovirus Sf9 system, the L-muCANPDELTA3 mutant lacks domain III, mutant L-muCANPDELTA4 lacks the calmodulin-like domain IV. In Sf9 cells co-expression of the inhibitor calpastatin is necessary to prevent autolysis of the L-muCANP subunit, whereas coexpression of the regulatory subunit enhances it. Only very low levels of mRNA of the truncated form L-muCANPDELTA4 are found in bacmid-transfectred Sf9 cells, and it proves impossible to isolate this mutant using the baculovirus expression system |
Homo sapiens |
Protein Variants
Protein Variants |
Comment |
Organism |
additional information |
the mutant enzyme L-muyCANPDELTA3 requires 0.4-0.53 mM of Ca2+ compared to 0.06 mM for the native enzyme, bacterially expressed mutant enzyme L-muyCANPDELTA3. A chimeric form composed of domains I-III of muCANP and domain IV of calpain II is also expressed in Sf9 cells. This mutant requires less Ca2+, 0.05 mM, than the native erythrocyte enzyme and has the highest specific activity of all calpains tested. All recombinant proteins are active as monomers in polyethylene glycol-containing buffers. The in vitro association with the regulatory subunit enhances only slightly the maximal velocity and the Ca2+ dependence of the expressed proteins |
Homo sapiens |
Metals/Ions
Metals/Ions |
Comment |
Organism |
Structure |
Ca2+ |
requires 0.053 mM Ca2+ for half-maximal activity. Activation by Ca2+ promotes the separation of the two subunits of the expressed recombinant protein |
Homo sapiens |
|
Organism
Organism |
UniProt |
Comment |
Textmining |
Homo sapiens |
- |
- |
- |
Renatured (Commentary)
Renatured (Comment) |
Organism |
presence of inhibitors during renaturation is necessary to prevent autolysis |
Homo sapiens |
Source Tissue
Source Tissue |
Comment |
Organism |
Textmining |
erythrocyte |
- |
Homo sapiens |
- |