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Literature summary extracted from

  • Filipek, R.; Rzychon, M.; Oleksy, A.; Gruca, M.; Dubin, A.; Potempa, J.; Bochtler, M.
    The staphostatin-staphopain complex: A forward binding inhibitor in complex with its target cysteine protease (2003), J. Biol. Chem., 278, 40959-40966.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.4.22.48 expression of large amounts of active wild-type enzyme in Escherichia coli is not possible, probably due to toxicity for the host Staphylococcus aureus
3.4.22.48 expression of wild-type enzyme as fusion protein in Escherichia coli, hybrid construct with a N-terminal GST-moiety linked to the proenzyme via a thrombin cleavable linker Staphylococcus aureus
3.4.22.48 overexpression of the inactive mutant in Escherichia coli BL21(DE3) as His-tagged protein Staphylococcus aureus

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.4.22.48 inactive mutant staphopain B in complex with inhibitor staphostatin B, 21°C, vapour diffusion method with sitting drops, reservoir solution: 2 M ammonium sulfate, 5% isopropanol, 0.8% v/v 1 M guanidinium hydrochloride, 1-2 weeks, cryoprotection in buffer containing a 9:1 mixture of 2.8 M ammonium sulfate and (2R,3R)-(-)-2,3-butanediol, injected into the drops, X-ray structure determination Staphylococcus aureus

Protein Variants

EC Number Protein Variants Comment Organism
3.4.22.48 additional information exchange of active site cysteine residue for an alanine, inactive mutant, fusion to N-terminal His-tag Staphylococcus aureus

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.4.22.48 staphostatin B endogenous inhibitor of cysteine proteases, recombinantly expressed in Escherichia coli as wild-type protein and mutant and purified; forms a mixed eight-stranded beta-barrel; structural interactions between inhibitor and enzyme are resolved from crystal structure of the enzyme-inhibitor complex Staphylococcus aureus

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
3.4.22.48 extracellular culture supernatant Staphylococcus aureus
-
-

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.4.22.48 peptide + H2O Staphylococcus aureus
-
 peptide proteolytically cleaved into fragments or single amino acids
-
 ?
3.4.22.48 protein + H2O Staphylococcus aureus
-
 protein proteolytically cleaved into peptide fragments
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
3.4.22.48 Staphylococcus aureus
-
 staphopain B
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.4.22.48 native from culture supernatant Staphylococcus aureus
3.4.22.48 recombinant inactive mutant form, His-tagged, from Escherichia coli Staphylococcus aureus
3.4.22.48 recombinant proenzyme form as GST-fusion protein from Escherichia coli, affinity chromatography, cleavage by V8 proteases to mature enzyme form Staphylococcus aureus

Reaction

EC Number Reaction Comment Organism Reaction ID
3.4.22.48 broad endopeptidase action on proteins including elastin, but rather limited hydrolysis of small-molecule substrates. Assays are conveniently made with hemoglobin, casein or Z-Phe-Arg-NHMec as substrate mechanism Staphylococcus aureus
a
3.4.22.48 broad endopeptidase action on proteins including elastin, but rather limited hydrolysis of small-molecule substrates. Assays are conveniently made with hemoglobin, casein or Z-Phe-Arg-NHMec as substrate active site structure Staphylococcus aureus
a

Source Tissue

EC Number Source Tissue Comment Organism Textmining
3.4.22.48 culture supernatant
-
 Staphylococcus aureus
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.4.22.48 peptide + H2O
-
 Staphylococcus aureus amino acids
-
 ?
3.4.22.48 peptide + H2O
-
 Staphylococcus aureus peptide proteolytically cleaved into fragments or single amino acids
-
 ?
3.4.22.48 protein + H2O
-
 Staphylococcus aureus peptide fragments
-
 ?
3.4.22.48 protein + H2O
-
 Staphylococcus aureus protein proteolytically cleaved into peptide fragments
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.4.22.48 staphopain B
-
 Staphylococcus aureus