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Literature summary extracted from

  • Wang, C.C.; Houng, H.C.; Chen, C.L.; Wang, P.J.; Kuo, C.F.; Lin, Y.S.; Wu, J.J.; Lin, M.T.; Liu, C.C.; Huang, W.; Chuang, W.J.
    Solution structure and backbone dynamics of Streptopain (2009), J. Biol. Chem., 284, 10957-10967.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.4.22.10 expressed in Escherichia coli Streptococcus pyogenes

Protein Variants

EC Number Protein Variants Comment Organism
3.4.22.10 C47S inactive mutant Streptococcus pyogenes
3.4.22.10 D9N protease activity (microgram/min/mg) substrate azocasein: 256, substrate proSPE B C47S mutant: 250, compared to wild-type 341 and 378, respectively Streptococcus pyogenes
3.4.22.10 G136A protease activity (microgram/min/mg) substrate azocasein: 22.3, substrate proSPE B C47S mutant: 8.1, compared to wild-type 341 and 378, respectively Streptococcus pyogenes
3.4.22.10 G163S protease activity (microgram/min/mg) substrate azocasein: 348, substrate proSPE B C47S mutant: 385, compared to wild-type 341 and 378, respectively Streptococcus pyogenes
3.4.22.10 G163S/A172S protease activity (microgram/min/mg) substrate azocasein: 286, substrate proSPE B C47S mutant: 236, compared to wild-type 341 and 378, respectively Streptococcus pyogenes
3.4.22.10 G239D protease activity (microgram/min/mg) substrate azocasein: 35.3, substrate proSPE B C47S mutant: 20, compared to wild-type 341 and 378, respectively Streptococcus pyogenes
3.4.22.10 V189A protease activity (microgram/min/mg) substrate azocasein: 2.6, substrate proSPE B C47S mutant: 0.9, compared to wild-type 341 and 378, respectively Streptococcus pyogenes
3.4.22.10 W212A protease activity (microgram/min/mg) substrate azocasein: 1.2, substrate proSPE B C47S mutant: 0.9, compared to wild-type 341 and 378, respectively Streptococcus pyogenes
3.4.22.10 W214A protease activity (microgram/min/mg) substrate azocasein: 0.8, substrate proSPE B C47S mutant: 0.8, compared to wild-type 341 and 378, respectively Streptococcus pyogenes

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.4.22.10 HgCl2 prevents the conversion to the mature 28 kDa form Streptococcus pyogenes
3.4.22.10 L-trans-epoxysuccinyl-leucylamido(4-guanidino)butane
-
 Streptococcus pyogenes

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.4.22.10 28000
-
 SDS-PAGE, mature form Streptococcus pyogenes
3.4.22.10 40000
-
 SDS-PAGE, zymogen Streptococcus pyogenes

Organism

EC Number Organism UniProt Comment Textmining
3.4.22.10 Streptococcus pyogenes
-
-
-

Posttranslational Modification

EC Number Posttranslational Modification Comment Organism
3.4.22.10 proteolytic modification SPE B produced from group A Streptococcus (GAS) is released extracellularly to culture medium as a zymogen (proSPE B) with a molecular mass of 40 kDa. The conversion of proSPE B to the 28 kDa active mature SPE B (mSPE B) is achieved by autoproteolysis and exogenous proteases Streptococcus pyogenes

Purification (Commentary)

EC Number Purification (Comment) Organism
3.4.22.10 the proteins are purified by Ni2+-chelating chromatography with a gradient of 20-200 mM imidazole. The proteins are concentrated by ultrafiltration using a 10 kDa cutoff membrane and then exchanged with phosphate-buffered saline Streptococcus pyogenes

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.4.22.10 341
-
 protease activity substrate azocasein Streptococcus pyogenes
3.4.22.10 378
-
 protease activity substrate proSPE B C47S mutant Streptococcus pyogenes

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.4.22.10 azocasein + H2O
-
 Streptococcus pyogenes ?
-
 ?
3.4.22.10 additional information the three-dimensional structure and backbone dynamics of the 28 kDa mature SPE B (mSPE B) is determined: Interactions between the C-terminal loop and the active site residues in mSPE B are observed. The structural differences between mSPE B and zymogen proSPE B are the conformation of the C-terminal loop and the orientation of the catalytic His-195 residue. Dynamics analysis of mSPEB and the mSPEB/inhibitor complexes show that the catalytic and C-terminal loops are the most flexible regions, suggesting that the flexible C-terminal loop of SPE B may play an important role in controlling the substrate binding, resulting in its broad substrate specificity Streptococcus pyogenes ?
-
 ?
3.4.22.10 proSPE B C47S + H2O the proSPE B C47S mutant is also be used as the substrate for activity assay because it does not exhibit any enzyme activity and exists as a 42 kDa zymogen Streptococcus pyogenes ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.4.22.10 Spe B
-
 Streptococcus pyogenes

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.4.22.10 37
-
 assay at Streptococcus pyogenes