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.

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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

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	-