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Literature summary extracted from

  • Liggieri, C.; Obregon, W.; Trejo, S.; Priolo, N.
    Biochemical analysis of a papain-like protease isolated from the latex of Asclepias curassavica L (2009), Acta Biochim. Biophys. Sin. (Shanghai), 41, 154-162.
    View publication on PubMed

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.4.22.7 (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
3.4.22.7 E-64
-
Asclepias curassavica
3.4.22.7 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
3.4.22.7 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]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.4.22.7 0.163
-
Nalpha-CBZ-L-Gln-4-nitrophenyl ester pH 8.5, 42°C Asclepias curassavica
3.4.22.7 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

EC Number Localization Comment Organism GeneOntology No. Textmining
3.4.22.7 extracellular asclepain cI Asclepias curassavica
-
-
3.4.22.7 extracellular asclepain cII Asclepias curassavica
-
-

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.4.22.7 23500
-
SDS-PAGE Asclepias curassavica
3.4.22.7 23500
-
x * 23500, asclepain cII, SDS-PAGE, x * 23590, asclepain cII, mass spectrometry Asclepias curassavica
3.4.22.7 23590
-
MALDI-TOF mass spectrometry Asclepias curassavica
3.4.22.7 23590
-
x * 23500, asclepain cII, SDS-PAGE, x * 23590, asclepain cII, mass spectrometry Asclepias curassavica

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.4.22.7 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 ?
-
?
3.4.22.7 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

EC Number Organism UniProt Comment Textmining
3.4.22.7 Asclepias curassavica
-
-
-
3.4.22.7 Asclepias curassavica B7VF64 fragment; scarlet milkweed
-
3.4.22.7 Asclepias curassavica B7VF65 fragment; scarlet milkweed
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.4.22.7 native asclepain cII 12.1fold to homogeneity by cation exchange chromatography, separation from asclepain cI Asclepias curassavica
3.4.22.7 separation of asclepain cI from asclepain cII Asclepias curassavica
3.4.22.7 SP-Sepharose column chromatography, gel filtration Asclepias curassavica

Source Tissue

EC Number Source Tissue Comment Organism Textmining
3.4.22.7 latex extracted from petioles Asclepias curassavica
-
3.4.22.7 latex latex extracted from petioles Asclepias curassavica
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.4.22.7 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
3.4.22.7 12.8
-
purified asclepain cII Asclepias curassavica

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.4.22.7 casein + H2O
-
Asclepias curassavica ?
-
?
3.4.22.7 L-pyroglutamyl-L-phenylalanyl-L-leucine 4-nitroanilide + H2O very low activity Asclepias curassavica L-pyroglutamyl-L-phenylalanyl-L-leucine + 4-nitroaniline
-
?
3.4.22.7 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 ?
-
?
3.4.22.7 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 ?
-
?
3.4.22.7 additional information the peptide PFLNA is a poor substrate for asclepain cII Asclepias curassavica ?
-
?
3.4.22.7 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 ?
-
?
3.4.22.7 N-alpha-benzyloxycarbonyl-Gly-4-nitrophenyl ester + H2O
-
Asclepias curassavica N-alpha-benzyloxycarbonyl-Gly + 4-nitrophenol
-
?
3.4.22.7 N-alpha-benzyloxycarbonyl-L-Ala-4-nitrophenyl ester + H2O
-
Asclepias curassavica N-alpha-benzyloxycarbonyl-L-Ala + 4-nitrophenol
-
?
3.4.22.7 N-alpha-benzyloxycarbonyl-L-Asn-4-nitrophenyl ester + H2O
-
Asclepias curassavica N-alpha-benzyloxycarbonyl-L-Asn + 4-nitrophenol
-
?
3.4.22.7 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
-
?
3.4.22.7 N-alpha-benzyloxycarbonyl-L-Gln-4-nitrophenyl ester + H2O
-
Asclepias curassavica N-alpha-benzyloxycarbonyl-L-Gln + 4-nitrophenol
-
?
3.4.22.7 N-alpha-benzyloxycarbonyl-L-Tyr-4-nitrophenyl ester + H2O
-
Asclepias curassavica N-alpha-benzyloxycarbonyl-L-Tyr + 4-nitrophenol
-
?
3.4.22.7 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

EC Number Subunits Comment Organism
3.4.22.7 ? x * 23500, asclepain cII, SDS-PAGE, x * 23590, asclepain cII, mass spectrometry Asclepias curassavica

Synonyms

EC Number Synonyms Comment Organism
3.4.22.7 asclepain cI
-
Asclepias curassavica
3.4.22.7 asclepain cII
-
Asclepias curassavica
3.4.22.7 papain-like endopeptidase
-
Asclepias curassavica
3.4.22.7 papain-like protease
-
Asclepias curassavica

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.4.22.7 42
-
assay at Asclepias curassavica

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.4.22.7 additional information
-
purified asclepain cII shows poor thermostability, profile, overview Asclepias curassavica
3.4.22.7 50
-
2 h, asclepain cII, 30% remaining activity Asclepias curassavica
3.4.22.7 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]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.4.22.7 121.5
-
Nalpha-CBZ-L-Gln-4-nitrophenyl ester pH 8.5, 42°C Asclepias curassavica
3.4.22.7 121.5
-
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

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.4.22.7 8.5
-
-
Asclepias curassavica
3.4.22.7 9.4 10.2 asclepain cII Asclepias curassavica

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.4.22.7 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

EC Number pH Stability pH Stability Maximum Comment Organism
3.4.22.7 10
-
1 h, asclepain cII, about 95% of the residual activity Asclepias curassavica
3.4.22.7 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

EC Number Organism Comment pI Value Maximum pI Value
3.4.22.7 Asclepias curassavica isoelectric focusing
-
9.3
3.4.22.7 Asclepias curassavica above, asclepain cII, isoelectric focusing
-
9.3

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.4.22.7 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