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Literature summary extracted from

  • Samanta, K.; Kar, P.; Chakraborti, T.; Shaikh, S.; Chakraborti, S.
    Characteristic properties of endoplasmic reticulum membrane m-calpain, calpastatin and lumen m-calpain: a comparative study between membrane and lumen m-calpains (2010), J. Biochem., 147, 765-779.
    View publication on PubMed

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.4.22.53 110 kDa calpastatin
-
 Bos taurus
3.4.22.53 70 kDa calpastatin
-
 Bos taurus
3.4.22.53 Calmidazolium calpain-specific inhibitor Bos taurus
3.4.22.53 leupeptin
-
 Bos taurus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.4.22.53 0.459
-
 succinyl-Leu-Leu-Val-Tyr-7-amido-4-methylcoumarin endoplasmic reticulum lumen m-calpain, in 100 mM imidazole-HCl buffer (pH 7.5), 5 mM L-cysteine, 2.5 mM 2-mercaptoethanol, 5 mM CaCl2, and 4% (v/v) dimethyl sulfoxide, at 30°C Bos taurus
3.4.22.53 0.461
-
 succinyl-Leu-Leu-Val-Tyr-7-amido-4-methylcoumarin endoplasmic reticulum membrane m-calpain, in 100 mM imidazole-HCl buffer (pH 7.5), 5 mM L-cysteine, 2.5 mM 2-mercaptoethanol, 5 mM CaCl2, and 4% (v/v) dimethyl sulfoxide, at 30°C Bos taurus
3.4.22.53 2.1
-
 succinyl-Leu-Tyr-7-amido-4-methylcoumarin endoplasmic reticulum lumen m-calpain, in 100 mM imidazole-HCl buffer (pH 7.5), 5 mM L-cysteine, 2.5 mM 2-mercaptoethanol, 5 mM CaCl2, and 4% (v/v) dimethyl sulfoxide, at 30°C Bos taurus
3.4.22.53 2.13
-
 succinyl-Leu-Tyr-7-amido-4-methylcoumarin endoplasmic reticulum membrane m-calpain, in 100 mM imidazole-HCl buffer (pH 7.5), 5 mM L-cysteine, 2.5 mM 2-mercaptoethanol, 5 mM CaCl2, and 4% (v/v) dimethyl sulfoxide, at 30°C Bos taurus
3.4.22.53 4.66
-
 succinyl-Leu-Met-7-amido-4-methylcoumarin endoplasmic reticulum membrane m-calpain, in 100 mM imidazole-HCl buffer (pH 7.5), 5 mM L-cysteine, 2.5 mM 2-mercaptoethanol, 5 mM CaCl2, and 4% (v/v) dimethyl sulfoxide, at 30°C Bos taurus
3.4.22.53 4.68
-
 succinyl-Leu-Met-7-amido-4-methylcoumarin endoplasmic reticulum lumen m-calpain, in 100 mM imidazole-HCl buffer (pH 7.5), 5 mM L-cysteine, 2.5 mM 2-mercaptoethanol, 5 mM CaCl2, and 4% (v/v) dimethyl sulfoxide, at 30°C Bos taurus
3.4.22.53 8.11
-
 t-butyloxycarbonyl-Val-Leu-Lys-7-amido-4-methylcoumarin endoplasmic reticulum membrane m-calpain, in 100 mM imidazole-HCl buffer (pH 7.5), 5 mM L-cysteine, 2.5 mM 2-mercaptoethanol, 5 mM CaCl2, and 4% (v/v) dimethyl sulfoxide, at 30°C Bos taurus
3.4.22.53 8.12
-
 t-butyloxycarbonyl-Val-Leu-Lys-7-amido-4-methylcoumarin endoplasmic reticulum lumen m-calpain, in 100 mM imidazole-HCl buffer (pH 7.5), 5 mM L-cysteine, 2.5 mM 2-mercaptoethanol, 5 mM CaCl2, and 4% (v/v) dimethyl sulfoxide, at 30°C Bos taurus

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
3.4.22.53 endoplasmic reticulum
-
 Bos taurus 5783
-

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.4.22.53 Ca2+ optimum activity at 5 mM Ca2+ Bos taurus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.4.22.53 8000
-
 1 * 8000 + 1 * 28000, gel filtration, the endoplasmic reticulum membrane m-calpain containing 80000 Da large and 28000 Da small subunit is non-phosphorylated Bos taurus
3.4.22.53 28000
-
 1 * 8000 + 1 * 28000, gel filtration, the endoplasmic reticulum membrane m-calpain containing 80000 Da large and 28000 Da small subunit is non-phosphorylated Bos taurus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.4.22.53 IP3R1 + H2O Bos taurus in presence of Ca2þ, m-calpain cleaves IP3R1 in the endoplasmic lumen ?
-
 ?
3.4.22.53 NCX1 + H2O Bos taurus
-
 ?
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
3.4.22.53 Bos taurus
-
-
-

Posttranslational Modification

EC Number Posttranslational Modification Comment Organism
3.4.22.53 phosphoprotein the endoplasmic reticulum lumen m-calpain containing only 80000 Da large subunit is phosphorylated Bos taurus

Purification (Commentary)

EC Number Purification (Comment) Organism
3.4.22.53 ammonium sulfate precipitation, DEAE-cellulose column chromatography, phenyl-Sepharose column chromatography, DEAE-TSK column chromatography, and Reactive Red gel filtration Bos taurus

Source Tissue

EC Number Source Tissue Comment Organism Textmining
3.4.22.53 pulmonary artery smooth muscle cell
-
 Bos taurus
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.4.22.53 additional information
-
 endoplasmic reticulum membrane m-calpain shows 170 units/mg specific activity after 323fold purification, endoplasmic reticulum lumen m-calpain shows 324 units/mg specific activity after 463fold purification one unit is defined as the amount of the enzyme that causes a change of 1.0 in A750 for the trichloroacetic acid-soluble products after incubation of the enzyme in the assay mixture Bos taurus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.4.22.53 casein + H2O
-
 Bos taurus ?
-
 ?
3.4.22.53 IP3R1 + H2O in presence of Ca2þ, m-calpain cleaves IP3R1 in the endoplasmic lumen Bos taurus ?
-
 ?
3.4.22.53 NCX1 + H2O
-
 Bos taurus ?
-
 ?
3.4.22.53 succinyl-Leu-Leu-Val-Tyr-7-amido-4-methylcoumarin + H2O
-
 Bos taurus succinyl-Leu-Leu-Val-Tyr + 7-amino-4-methylcoumarin
-
 ?
3.4.22.53 succinyl-Leu-Met-7-amido-4-methylcoumarin + H2O
-
 Bos taurus succinyl-Leu-Met + 7-amino-4-methylcoumarin
-
 ?
3.4.22.53 succinyl-Leu-Tyr-7-amido-4-methylcoumarin + H2O
-
 Bos taurus succinyl-Leu-Tyr + 7-amino-4-methylcoumarin
-
 ?
3.4.22.53 t-butyloxycarbonyl-Val-Leu-Lys-7-amido-4-methylcoumarin + H2O
-
 Bos taurus t-butyloxycarbonyl-Val-Leu-Lys + 7-amino-4-methylcoumarin
-
 ?

Subunits

EC Number Subunits Comment Organism
3.4.22.53 heterodimer 1 * 8000 + 1 * 28000, gel filtration, the endoplasmic reticulum membrane m-calpain containing 80000 Da large and 28000 Da small subunit is non-phosphorylated Bos taurus

Synonyms

EC Number Synonyms Comment Organism
3.4.22.53 m-calpain
-
 Bos taurus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.4.22.53 0.062
-
 succinyl-Leu-Leu-Val-Tyr-7-amido-4-methylcoumarin endoplasmic reticulum lumen m-calpain, in 100 mM imidazole-HCl buffer (pH 7.5), 5 mM L-cysteine, 2.5 mM 2-mercaptoethanol, 5 mM CaCl2, and 4% (v/v) dimethyl sulfoxide, at 30°C Bos taurus
3.4.22.53 0.063
-
 succinyl-Leu-Leu-Val-Tyr-7-amido-4-methylcoumarin endoplasmic reticulum membrane m-calpain, in 100 mM imidazole-HCl buffer (pH 7.5), 5 mM L-cysteine, 2.5 mM 2-mercaptoethanol, 5 mM CaCl2, and 4% (v/v) dimethyl sulfoxide, at 30°C Bos taurus
3.4.22.53 0.083
-
 succinyl-Leu-Tyr-7-amido-4-methylcoumarin endoplasmic reticulum lumen m-calpain, in 100 mM imidazole-HCl buffer (pH 7.5), 5 mM L-cysteine, 2.5 mM 2-mercaptoethanol, 5 mM CaCl2, and 4% (v/v) dimethyl sulfoxide, at 30°C Bos taurus
3.4.22.53 0.084
-
 succinyl-Leu-Tyr-7-amido-4-methylcoumarin endoplasmic reticulum membrane m-calpain, in 100 mM imidazole-HCl buffer (pH 7.5), 5 mM L-cysteine, 2.5 mM 2-mercaptoethanol, 5 mM CaCl2, and 4% (v/v) dimethyl sulfoxide, at 30°C Bos taurus
3.4.22.53 0.186
-
 succinyl-Leu-Met-7-amido-4-methylcoumarin endoplasmic reticulum membrane m-calpain, in 100 mM imidazole-HCl buffer (pH 7.5), 5 mM L-cysteine, 2.5 mM 2-mercaptoethanol, 5 mM CaCl2, and 4% (v/v) dimethyl sulfoxide, at 30°C Bos taurus
3.4.22.53 0.189
-
 succinyl-Leu-Met-7-amido-4-methylcoumarin endoplasmic reticulum lumen m-calpain, in 100 mM imidazole-HCl buffer (pH 7.5), 5 mM L-cysteine, 2.5 mM 2-mercaptoethanol, 5 mM CaCl2, and 4% (v/v) dimethyl sulfoxide, at 30°C Bos taurus
3.4.22.53 1.06
-
 t-butyloxycarbonyl-Val-Leu-Lys-7-amido-4-methylcoumarin endoplasmic reticulum membrane m-calpain, in 100 mM imidazole-HCl buffer (pH 7.5), 5 mM L-cysteine, 2.5 mM 2-mercaptoethanol, 5 mM CaCl2, and 4% (v/v) dimethyl sulfoxide, at 30°C Bos taurus
3.4.22.53 1.08
-
 t-butyloxycarbonyl-Val-Leu-Lys-7-amido-4-methylcoumarin endoplasmic reticulum lumen m-calpain, in 100 mM imidazole-HCl buffer (pH 7.5), 5 mM L-cysteine, 2.5 mM 2-mercaptoethanol, 5 mM CaCl2, and 4% (v/v) dimethyl sulfoxide, at 30°C Bos taurus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.4.22.53 7.5
-
-
 Bos taurus

pI Value

EC Number Organism Comment pI Value Maximum pI Value
3.4.22.53 Bos taurus isoelectric focusing
-
 4.5

IC50 Value

EC Number IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
3.4.22.53 0.00000052
-
 in 100 mM imidazole-HCl buffer (pH 7.5),5 mM cysteine and 5 mM CaCl2, at 30°C Bos taurus 110 kDa calpastatin
3.4.22.53 0.0000008
-
 in 100 mM imidazole-HCl buffer (pH 7.5),5 mM cysteine and 5 mM CaCl2, at 30°C Bos taurus 70 kDa calpastatin

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.4.22.53 0.0394
-
 succinyl-Leu-Tyr-7-amido-4-methylcoumarin endoplasmic reticulum membrane m-calpain, in 100 mM imidazole-HCl buffer (pH 7.5), 5 mM L-cysteine, 2.5 mM 2-mercaptoethanol, 5 mM CaCl2, and 4% (v/v) dimethyl sulfoxide, at 30°C Bos taurus
3.4.22.53 0.0395
-
 succinyl-Leu-Tyr-7-amido-4-methylcoumarin endoplasmic reticulum lumen m-calpain, in 100 mM imidazole-HCl buffer (pH 7.5), 5 mM L-cysteine, 2.5 mM 2-mercaptoethanol, 5 mM CaCl2, and 4% (v/v) dimethyl sulfoxide, at 30°C Bos taurus
3.4.22.53 0.0399
-
 succinyl-Leu-Met-7-amido-4-methylcoumarin endoplasmic reticulum membrane m-calpain, in 100 mM imidazole-HCl buffer (pH 7.5), 5 mM L-cysteine, 2.5 mM 2-mercaptoethanol, 5 mM CaCl2, and 4% (v/v) dimethyl sulfoxide, at 30°C Bos taurus
3.4.22.53 0.0404
-
 succinyl-Leu-Met-7-amido-4-methylcoumarin endoplasmic reticulum lumen m-calpain, in 100 mM imidazole-HCl buffer (pH 7.5), 5 mM L-cysteine, 2.5 mM 2-mercaptoethanol, 5 mM CaCl2, and 4% (v/v) dimethyl sulfoxide, at 30°C Bos taurus
3.4.22.53 0.131
-
 t-butyloxycarbonyl-Val-Leu-Lys-7-amido-4-methylcoumarin endoplasmic reticulum membrane m-calpain, in 100 mM imidazole-HCl buffer (pH 7.5), 5 mM L-cysteine, 2.5 mM 2-mercaptoethanol, 5 mM CaCl2, and 4% (v/v) dimethyl sulfoxide, at 30°C Bos taurus
3.4.22.53 0.133
-
 t-butyloxycarbonyl-Val-Leu-Lys-7-amido-4-methylcoumarin endoplasmic reticulum lumen m-calpain, in 100 mM imidazole-HCl buffer (pH 7.5), 5 mM L-cysteine, 2.5 mM 2-mercaptoethanol, 5 mM CaCl2, and 4% (v/v) dimethyl sulfoxide, at 30°C Bos taurus
3.4.22.53 0.135
-
 succinyl-Leu-Leu-Val-Tyr-7-amido-4-methylcoumarin endoplasmic reticulum lumen m-calpain, in 100 mM imidazole-HCl buffer (pH 7.5), 5 mM L-cysteine, 2.5 mM 2-mercaptoethanol, 5 mM CaCl2, and 4% (v/v) dimethyl sulfoxide, at 30°C Bos taurus
3.4.22.53 0.137
-
 succinyl-Leu-Leu-Val-Tyr-7-amido-4-methylcoumarin endoplasmic reticulum membrane m-calpain, in 100 mM imidazole-HCl buffer (pH 7.5), 5 mM L-cysteine, 2.5 mM 2-mercaptoethanol, 5 mM CaCl2, and 4% (v/v) dimethyl sulfoxide, at 30°C Bos taurus