Literature summary for 3.4.13.9 extracted from

Maher, M.J.; Ghosh, M.; Grunden, A.M.; Menon, A.L.; Adams, M.W.; Freeman, H.C.; Guss, J.M.
Structure of the prolidase from Pyrococcus furiosus (2004), Biochemistry, 43, 2771-2783.

Crystallization (Commentary)

Crystallization (Comment)	Organism
0.0025 ml of purified recombinant enzyme, 10 mg/ml, in 50 mM Tris-HCl, pH 8.5, 6.5-7.5% PEG 8000, 0.1 M magnesium acetate, hanging drop vapour diffusion, against 1.0 ml reservoir solution containing 13-15% PEG 8000, 0.2 M magnesium acetate, 50 mM Tris-HCl, pH 8.5, 20°C, 180 days, addition of solid (2S,3R)-3-amino-2-hydroxy-5-methyl-hexanoyl-proline and 15% 2-methyl-2,4-pentanediol for formation and crystallization of enzyme-inhibitor complex, X-ray structure determination and analysis at 2.0 A resolution of crystal form II	Pyrococcus furiosus
native enzyme and in complex with inhibitor (2S,3R)-3-amino-2-hydroxy-5-methylhexanoyl-proline. One homodimer per asymmetric unit. Each subunit has two domains, the C-terminal domain includes the catalytic site centered on a dinuclear metal cluster	Pyrococcus furiosus

Crystallization (Comment)

Organism

0.0025 ml of purified recombinant enzyme, 10 mg/ml, in 50 mM Tris-HCl, pH 8.5, 6.5-7.5% PEG 8000, 0.1 M magnesium acetate, hanging drop vapour diffusion, against 1.0 ml reservoir solution containing 13-15% PEG 8000, 0.2 M magnesium acetate, 50 mM Tris-HCl, pH 8.5, 20°C, 180 days, addition of solid (2S,3R)-3-amino-2-hydroxy-5-methyl-hexanoyl-proline and 15% 2-methyl-2,4-pentanediol for formation and crystallization of enzyme-inhibitor complex, X-ray structure determination and analysis at 2.0 A resolution of crystal form II

Pyrococcus furiosus

native enzyme and in complex with inhibitor (2S,3R)-3-amino-2-hydroxy-5-methylhexanoyl-proline. One homodimer per asymmetric unit. Each subunit has two domains, the C-terminal domain includes the catalytic site centered on a dinuclear metal cluster

Pyrococcus furiosus

Inhibitors

Inhibitors	Comment	Organism	Structure
(2S,3R)-3-amino-2-hydroxy-5-methyl-hexanoyl-proline	-	Pyrococcus furiosus
(2S,3R)-3-amino-2-hydroxy-5-methylhexanoyl-proline	-	Pyrococcus furiosus

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Co2+	dinuclear metal cluster in the active site, binding of 2 Co2+ per subunit	Pyrococcus furiosus
Zn2+	2 Zn2+ bound to the subunit in the crystallized enzyme only replacing the Co2+ ions, binding structure via 5 coordinates	Pyrococcus furiosus

Organism

Organism	UniProt	Comment	Textmining
Pyrococcus furiosus	-	-	-
Pyrococcus furiosus	P81535	-	-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
590	-	purified crystallized recombinant enzyme	Pyrococcus furiosus
1300	-	purified noncrystallized recombinant enzyme	Pyrococcus furiosus

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
Xaa-Pro + H2O	-	Pyrococcus furiosus	Xaa + Pro	-	?

Subunits

Subunits	Comment	Organism
dimer	crystal structure, 2 subunits each with 2 domains, with the C-terminal domain bearing the active site	Pyrococcus furiosus

Synonyms

Synonyms	Comment	Organism
More	enzyme belongs to the methionyl amino peptidase family M24, fomerly EC 3.4.3.7	Pyrococcus furiosus
Pfprol	-	Pyrococcus furiosus
prolidase	-	Pyrococcus furiosus

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
100	-	assay at	Pyrococcus furiosus