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Literature summary for 3.4.22.7 extracted from
- Biochemical analysis of a papain-like protease isolated from the latex of Asclepias curassavica L (2009), Acta Biochim. Biophys. Sin. (Shanghai), 41, 154-162.
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| (2S,3S)-3-(N-[(S)-1-[N-(4-guanidinobutyl)carbamoyl]3-methylbutyl]carbamoyl)oxirane-2-carboxylic acid | i.e. E-64, complete immediate and irreversible inhibition at 10 mM after 30 min incubation | Asclepias curassavica |  |
| E-64 | - |
Asclepias curassavica | |
| additional information | 1,10-phenantroline and pepstatine do not affect the activity of asclepain cII; activity of asclepain cII is inhibited by cysteine proteases inhibitors like E-64, but not by any other protease inhibitors such as 1,10-phenantroline, phenylmethanesulfonyl fluoride, and pepstatine | Asclepias curassavica | |
| phenylmethylsulfonyl fluoride | incubation with 100 mM phenylmethylsulfonyl fluoride for 30 min decreases activity to 63% of the initial value, but inhibition is partially reverted when 12 mM cysteine is added to the reaction mixture | Asclepias curassavica | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.163 | - |
Nalpha-CBZ-L-Gln-4-nitrophenyl ester | pH 8.5, 42°C | Asclepias curassavica | |
| 0.1634 | - |
N-alpha-benzyloxycarbonyl-L-Gln-4-nitrophenyl ester | at 37°C, in 0.1 M Tris-HCl buffer (pH 8.0) containing 2 mM EDTA and 25 mM cysteine | Asclepias curassavica | |
Localization
| Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|
| extracellular | asclepain cI | Asclepias curassavica | - |
- |
| extracellular | asclepain cII | Asclepias curassavica | - |
- |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 23500 | - |
SDS-PAGE | Asclepias curassavica |
| 23500 | - |
x * 23500, asclepain cII, SDS-PAGE, x * 23590, asclepain cII, mass spectrometry | Asclepias curassavica |
| 23590 | - |
MALDI-TOF mass spectrometry | Asclepias curassavica |
| 23590 | - |
x * 23500, asclepain cII, SDS-PAGE, x * 23590, asclepain cII, mass spectrometry | Asclepias curassavica |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| additional information | Asclepias curassavica | asclepain cI is the main active enzyme, but shows lower specific activity than asclepain cII, which is the minor proteolytic component in the latex, overview | ? | - |
? | |
| additional information | Asclepias curassavica | asclepain cII is the minor proteolytic component in the latex, but shows higher specific activity than asclepain cI, the main active enzyme, overview | ? | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Asclepias curassavica | - |
- |
- |
| Asclepias curassavica | B7VF64 | fragment; scarlet milkweed | - |
| Asclepias curassavica | B7VF65 | fragment; scarlet milkweed | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| native asclepain cII 12.1fold to homogeneity by cation exchange chromatography, separation from asclepain cI | Asclepias curassavica |
| separation of asclepain cI from asclepain cII | Asclepias curassavica |
| SP-Sepharose column chromatography, gel filtration | Asclepias curassavica |
Source Tissue
| Source Tissue | Comment | Organism | Textmining |
|---|---|---|---|
| latex | extracted from petioles | Asclepias curassavica | - |
| latex | latex extracted from petioles | Asclepias curassavica | - |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| additional information | - |
the enzyme has a specific activity of 1.0586 units/mg in crude extract and of 12.836 units/mg after 12.1 fold purification, using casein as substrate, one arbitrary enzyme unit is defined as the amount of enzyme that produces an increase of one absorbance unit per minute | Asclepias curassavica |
| 12.8 | - |
purified asclepain cII | Asclepias curassavica |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| casein + H2O | - |
Asclepias curassavica | ? | - |
? | |
| L-pyroglutamyl-L-phenylalanyl-L-leucine 4-nitroanilide + H2O | very low activity | Asclepias curassavica | L-pyroglutamyl-L-phenylalanyl-L-leucine + 4-nitroaniline | - |
? | |
| additional information | asclepain cI is the main active enzyme, but shows lower specific activity than asclepain cII, which is the minor proteolytic component in the latex, overview | Asclepias curassavica | ? | - |
? | |
| additional information | asclepain cII is the minor proteolytic component in the latex, but shows higher specific activity than asclepain cI, the main active enzyme, overview | Asclepias curassavica | ? | - |
? | |
| additional information | the peptide PFLNA is a poor substrate for asclepain cII | Asclepias curassavica | ? | - |
? | |
| additional information | no activity towards N-alpha-benzyloxycarbonyl-L-Val-4-nitrophenyl ester, N-alpha-benzyloxycarbonyl-L-Leu-4-nitrophenyl ester, N-alpha-benzyloxycarbonyl-L-Ile-4-nitrophenyl ester, and N-alpha-benzyloxycarbonyl-L-Lys-4-nitrophenyl ester | Asclepias curassavica | ? | - |
? | |
| N-alpha-benzyloxycarbonyl-Gly-4-nitrophenyl ester + H2O | - |
Asclepias curassavica | N-alpha-benzyloxycarbonyl-Gly + 4-nitrophenol | - |
? | |
| N-alpha-benzyloxycarbonyl-L-Ala-4-nitrophenyl ester + H2O | - |
Asclepias curassavica | N-alpha-benzyloxycarbonyl-L-Ala + 4-nitrophenol | - |
? | |
| N-alpha-benzyloxycarbonyl-L-Asn-4-nitrophenyl ester + H2O | - |
Asclepias curassavica | N-alpha-benzyloxycarbonyl-L-Asn + 4-nitrophenol | - |
? | |
| N-alpha-benzyloxycarbonyl-L-Asp-4-nitrophenyl ester + H2O | asclepain cII has highest endoesterolytic activity using N-alpha-benzyloxycarbonyl-amino acid p-nitrophenyl esters with L-Asp as amino acid | Asclepias curassavica | N-alpha-benzyloxycarbonyl-L-Asp + 4-nitrophenol | - |
? | |
| N-alpha-benzyloxycarbonyl-L-Gln-4-nitrophenyl ester + H2O | - |
Asclepias curassavica | N-alpha-benzyloxycarbonyl-L-Gln + 4-nitrophenol | - |
? | |
| N-alpha-benzyloxycarbonyl-L-Tyr-4-nitrophenyl ester + H2O | - |
Asclepias curassavica | N-alpha-benzyloxycarbonyl-L-Tyr + 4-nitrophenol | - |
? | |
| Nalpha-CBZ-L-Gln-4-nitrophenyl ester + H2O | when assayed on N-alpha-CBZ-amino acid-4-nitrophenyl esters, the enzyme exhibits higher preference for the glutamine derivative | Asclepias curassavica | Nalpha-CBZ-L-Gln + 4-nitrophenol | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| ? | x * 23500, asclepain cII, SDS-PAGE, x * 23590, asclepain cII, mass spectrometry | Asclepias curassavica |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| asclepain cI | - |
Asclepias curassavica |
| asclepain cII | - |
Asclepias curassavica |
| papain-like endopeptidase | - |
Asclepias curassavica |
| papain-like protease | - |
Asclepias curassavica |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 42 | - |
assay at | Asclepias curassavica |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| additional information | - |
purified asclepain cII shows poor thermostability, profile, overview | Asclepias curassavica |
| 50 | - |
2 h, asclepain cII, 30% remaining activity | Asclepias curassavica |
| 50 | 70 | asclepain cII exhibits poor thermostability, as only 30% of its activity is retained after 2 h of incubation at 50°C, at 60-70°C the loss of activity is rapid | Asclepias curassavica |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 121.5 | - |
Nalpha-CBZ-L-Gln-4-nitrophenyl ester | pH 8.5, 42°C | Asclepias curassavica | |
| 121.5 | - |
N-alpha-benzyloxycarbonyl-L-Gln-4-nitrophenyl ester | at 37°C, in 0.1 M Tris-HCl buffer (pH 8.0) containing 2 mM EDTA and 25 mM cysteine | Asclepias curassavica | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 8.5 | - |
- |
Asclepias curassavica |
| 9.4 | 10.2 | asclepain cII | Asclepias curassavica |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 9.4 | 10.2 | asclepain cII shows maximum activity within the range of pH 9.4 and 10.2 with casein as substrate | Asclepias curassavica |
pH Stability
| pH Stability | pH Stability Maximum | Comment | Organism |
|---|---|---|---|
| 10 | - |
1 h, asclepain cII, about 95% of the residual activity | Asclepias curassavica |
| 10 | - |
asclepain cII shows a high stability at pH 10.0 retaining about 95% of the residual activity after 1 h | Asclepias curassavica |
pI Value
| Organism | Comment | pI Value Maximum | pI Value |
|---|---|---|---|
| Asclepias curassavica | isoelectric focusing | - |
9.3 |
| Asclepias curassavica | above, asclepain cII, isoelectric focusing | - |
9.3 |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 740000 | - |
N-alpha-benzyloxycarbonyl-L-Gln-4-nitrophenyl ester | at 37°C, in 0.1 M Tris-HCl buffer (pH 8.0) containing 2 mM EDTA and 25 mM cysteine | Asclepias curassavica | |
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