Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 3.4.21.62 extracted from

  • Pulido, M.A.; Tanaka, S.; Sringiew, C.; You, D.J.; Matsumura, H.; Koga, Y.; Takano, K.; Kanaya, S.
    Requirement of left-handed glycine residue for high stability of the Tk-subtilisin propeptide as revealed by mutational and crystallographic analyses (2007), J. Mol. Biol., 374, 1359-1373.
    View publication on PubMed

Application

Application Comment Organism
additional information Gly56 is required in stabilizing the propeptide fold. Stabilization of this fold leads to strong binding of the propeptide to subtilisin, high resistance of the propeptide to proteolytic degradation, and slow maturation of Pro-subtilisin Thermococcus kodakarensis

Cloned(Commentary)

Cloned (Comment) Organism
pET25b derivatives for overproduction of Pro-G56W, Pro-G56E, G56W-propeptide, and G56E-propeptide. G56W-propeptide overproduced in soluble form, while G56E-propeptide overproduced in insoluble form. S324A-subtilisin overproduced in inclusion bodies Thermococcus kodakarensis

Crystallization (Commentary)

Crystallization (Comment) Organism
by the sitting drop vapor diffusion method. Crystal structures of propeptides in complex with S324A-subtilisin. Conformation of the propeptide is altered by the mutation, such that nonglycine residues at position 56 assume a right-handed conformation and hydrophobic interactions at the core region decrease Thermococcus kodakarensis

Protein Variants

Protein Variants Comment Organism
G56E Pro-G56E gives a halo at temperatures equal or higher than 60 °C Thermococcus kodakarensis
G56F Pro-G56F gives a halo only at temperatures equal or higher than 70 °C Thermococcus kodakarensis
G56S Pro-G56S gives a halo from 40-80°C Thermococcus kodakarensis
G56W Pro-G56W gives a halo only at 80 °C like the wild-type Thermococcus kodakarensis
S324A active-site mutant of Pro-subtilisin Thermococcus kodakarensis

Inhibitors

Inhibitors Comment Organism Structure
propeptide inhibition in a concentration-dependent manner. Wild-type propeptide is more potent than G56W-, G56S- and G56E-propeptide Thermococcus kodakarensis

Metals/Ions

Metals/Ions Comment Organism Structure
Ca2+
-
Thermococcus kodakarensis

Organism

Organism UniProt Comment Textmining
Thermococcus kodakarensis
-
-
-

Purification (Commentary)

Purification (Comment) Organism
G56W-propeptide and G56E-propeptide, S324A-subtilisin purified in denatured form Thermococcus kodakarensis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
casein + H2O
-
Thermococcus kodakarensis ?
-
?
N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide + H2O
-
Thermococcus kodakarensis N-succinyl-Ala-Ala-Pro-Phe + p-nitroaniline
-
?

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
40 80
-
Thermococcus kodakarensis