Literature summary for 3.4.22.48 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.

Search for more literature for 3.4.22.48

Cloned(Commentary)

Cloned (Comment)	Organism
expression of large amounts of active wild-type enzyme in Escherichia coli is not possible, probably due to toxicity for the host	Staphylococcus aureus
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
overexpression of the inactive mutant in Escherichia coli BL21(DE3) as His-tagged protein	Staphylococcus aureus

Crystallization (Commentary)

Crystallization (Comment)	Organism
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

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

Inhibitors

Inhibitors	Comment	Organism	Structure
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

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

Natural Substrates/ Products (Substrates)

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

Organism

Organism	UniProt	Comment	Textmining
Staphylococcus aureus	-	staphopain B	-

Purification (Commentary)

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

Reaction

Reaction	Comment	Organism	Reaction ID
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
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

Source Tissue	Comment	Organism	Textmining
culture supernatant	-	Staphylococcus aureus	-

Substrates and Products (Substrate)

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

Synonyms

Synonyms	Comment	Organism
staphopain B	-	Staphylococcus aureus

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Release 2023.1 (January 2023)