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Literature summary extracted from

  • Mihelic, M.; Dobersek, A.; Guncar, G.; Turk, D.
    Inhibitory fragment from the p41 form of invariant chain can regulate activity of cysteine cathepsins in antigen presentation (2008), J. Biol. Chem., 283, 14453-14460.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.4.22.15 amplification of cDNA by PCR, PCR-products cloned into Pichia pastoris expression vector pPIC9K with His-tag, transformation of Pichia pastoris strain GS115, 4 days of expression Homo sapiens
3.4.22.15 amplification of cDNA by PCR, PCR-products cloned into Pichia pastoris expression vector pPIC9K with His-tag, transformation of Pichia pastoris strain GS115, 4 days of expression Mus musculus

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.4.22.15 modelling 3D-structure of cathepsin L (PDB entry 1ICF), analysing residues binding the inhibitor p41-fragment, comparing binding sites of different Cathepsins Homo sapiens
3.4.22.15 modelling 3D-structure of cathepsin L (PDB entry 1ICF), analysing residues binding the inhibitor p41-fragment, comparing binding sites of different Cathepsins Mus musculus
3.4.22.15 molecular modeling of complex with fragment p41 of major histocompatibility complex class II-associated invariant chain Mus musculus
3.4.22.15 molecular modeling of complex with fragment p41 of major histocompatibility complex class II-associated invariant chain Homo sapiens
3.4.22.43 molecular modeling of complex with fragment p41 of major histocompatibility complex class II-associated invariant chain Homo sapiens

Protein Variants

EC Number Protein Variants Comment Organism
3.4.22.15 G139R 2fold decrease in binding affinity to the inhibitory fragment p41 of major histocompatibility complex class II-associated invariant chain Homo sapiens
3.4.22.15 G139R KI-value increased compared to wild-type Homo sapiens

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.4.22.15 fragment p41 of major histocompatibility complex class II-associated invariant chain inhibitory to human cathepsin V, cathepsin L, cathepsin K, cathepsin F with Ki values in the low nanomolar range. Ki values are sufficiently low to ensure complex formation at physiological concentrations. Regulation of the proteolytic activity of most of the cysteine cathepsins by the p41 fragment is an important and widespread control mechanism of antigen presentation Homo sapiens
3.4.22.15 fragment p41 of major histocompatibility complex class II-associated invariant chain inhibitory to human cathepsin V, cathepsin L, cathepsin K, cathepsin F with Ki values in the low nanomolar range. Ki values are sufficiently low to ensure complex formation at physiological concentrations. Regulation of the proteolytic activity of most of the cysteine cathepsins by the p41 fragment is an important and widespread control mechanism of antigen presentation Mus musculus
3.4.22.15 p41-fragment-human 64 residues present in the p41 form of the human major histocompatibility complex II (MHCII)-associated invariant chain Homo sapiens
3.4.22.15 p41-fragment-human 64 residues present in the p41 form of the human major histocompatibility complex II (MHCII)-associated invariant chain Mus musculus
3.4.22.15 p41-fragment-mouse 64 residues present in the p41 form of the murine major histocompatibility complex II (MHCII)-associated invariant chain Mus musculus
3.4.22.43 fragment p41 of major histocompatibility complex class II-associated invariant chain inhibitory to human cathepsin V, cathepsin L, cathepsin K, cathepsin F with Ki values in the low nanomolar range. Ki values are sufficiently low to ensure complex formation at physiological concentrations. Regulation of the proteolytic activity of most of the cysteine cathepsins by the p41 fragment is an important and widespread control mechanism of antigen presentation Homo sapiens

Organism

EC Number Organism UniProt Comment Textmining
3.4.22.15 Homo sapiens
-
-
-
3.4.22.15 Homo sapiens P07711
-
-
3.4.22.15 Mus musculus
-
-
-
3.4.22.15 Mus musculus P06797
-
-
3.4.22.43 Homo sapiens
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.4.22.15 Ni-Sepharose 6 Fast Flow column, eluted with binding buffer containing 500 mM imidazole, activation of immature cathepsin L by dialysis, Sephadex-75 HR column, dialysis, blocked with 10-fold molar excess of methyl-methanethiosulfonate Homo sapiens
3.4.22.15 Ni-Sepharose 6 Fast Flow column, eluted with binding buffer containing 500 mM imidazole, activation of immature cathepsin L by dialysis, Sephadex-75 HR column, dialysis, blocked with 10-fold molar excess of methyl-methanethiosulfonate Mus musculus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.4.22.15 benzyloxycarbonyl-Phe-Arg-4-methylcoumarin-7-amide + H2O
-
 Homo sapiens benzyloxycarbonyl-Phe-Arg + 7-amino-4-methylcoumarin
-
 ?
3.4.22.15 benzyloxycarbonyl-Phe-Arg-4-methylcoumarin-7-amide + H2O
-
 Mus musculus benzyloxycarbonyl-Phe-Arg + 7-amino-4-methylcoumarin
-
 ?
3.4.22.15 benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin + H2O
-
 Mus musculus benzyloxycarbonyl-Phe-Arg + 7-amino-4-methylcoumarin
-
 ?
3.4.22.15 benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin + H2O
-
 Homo sapiens benzyloxycarbonyl-Phe-Arg + 7-amino-4-methylcoumarin
-
 ?
3.4.22.43 benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin + H2O
-
 Homo sapiens benzyloxycarbonyl-Phe-Arg + 7-amino-4-methylcoumarin
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.4.22.15 cathepsin L
-
 Homo sapiens
3.4.22.15 cathepsin L
-
 Mus musculus

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
3.4.22.15 0.00000000149
-
 p41-fragment-human substrate: benzyloxycarbonyl-Phe-Arg-4-methylcoumarin-7-amide, production rate of 7-amino-4-methylcoumarin measured, excitation and emission wavelengths of 370 and 460 nm, pseudo first order reaction conditions (inhibitor concentration at least 10-fold higher than enzyme concentration) Mus musculus
3.4.22.15 0.00000000552
-
 p41-fragment-human wild-type, substrate: benzyloxycarbonyl-Phe-Arg-4-methylcoumarin-7-amide, production rate of 7-amino-4-methylcoumarin measured, excitation and emission wavelengths of 370 and 460 nm, pseudo first order reaction conditions (inhibitor concentration at least 10-fold higher than enzyme concentration) Homo sapiens
3.4.22.15 0.0000000072
-
 fragment p41 of major histocompatibility complex class II-associated invariant chain pH 6.0, 25°C Mus musculus
3.4.22.15 0.00000000722
-
 p41-fragment-mouse substrate: benzyloxycarbonyl-Phe-Arg-4-methylcoumarin-7-amide, production rate of 7-amino-4-methylcoumarin measured, excitation and emission wavelengths of 370 and 460 nm, pseudo first order reaction conditions (inhibitor concentration at least 10-fold higher than enzyme concentration) Mus musculus
3.4.22.15 0.0000000101
-
 fragment p41 of major histocompatibility complex class II-associated invariant chain pH 6.0, 25°C Homo sapiens
3.4.22.15 0.0000000101
-
 p41-fragment-human G139R mutant, substrate: benzyloxycarbonyl-Phe-Arg-4-methylcoumarin-7-amide, production rate of 7-amino-4-methylcoumarin measured, excitation and emission wavelengths of 370 and 460 nm, pseudo first order reaction conditions (inhibitor concentration at least 10-fold higher than enzyme concentration) Homo sapiens
3.4.22.43 0.0000000072
-
 fragment p41 of major histocompatibility complex class II-associated invariant chain pH 6.0, 25°C Homo sapiens