Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli | Streptococcus pyogenes |
Protein Variants | Comment | Organism |
---|---|---|
C47S | inactive mutant | Streptococcus pyogenes |
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 |
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 |
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 |
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 |
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 |
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 |
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 |
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 | Comment | Organism | Structure |
---|---|---|---|
HgCl2 | prevents the conversion to the mature 28 kDa form | Streptococcus pyogenes | |
L-trans-epoxysuccinyl-leucylamido(4-guanidino)butane | - |
Streptococcus pyogenes |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
28000 | - |
SDS-PAGE, mature form | Streptococcus pyogenes |
40000 | - |
SDS-PAGE, zymogen | Streptococcus pyogenes |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Streptococcus pyogenes | - |
- |
- |
Posttranslational Modification | Comment | Organism |
---|---|---|
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 (Comment) | Organism |
---|---|
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 Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
341 | - |
protease activity substrate azocasein | Streptococcus pyogenes |
378 | - |
protease activity substrate proSPE B C47S mutant | Streptococcus pyogenes |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
azocasein + H2O | - |
Streptococcus pyogenes | ? | - |
? | |
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 | ? | - |
? | |
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 | Comment | Organism |
---|---|---|
Spe B | - |
Streptococcus pyogenes |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Streptococcus pyogenes |