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Literature summary extracted from

  • Tanaka, S.; Saito, K.; Chon, H.; Matsumura, H.; Koga, Y.; Takano, K.; Kanaya, S.
    Crystallization and preliminary X-ray diffraction study of an active-site mutant of pro-Tk-subtilisin from a hyperthermophilic archaeon (2006), Acta Crystallogr. Sect. F, 62, 902-905.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.4.21.B57 overproduction of pro-S255A in Escherichia coli BL21-Codon-Plus(DE3) Thermococcus kodakarensis
3.4.21.62 overexpression of pro-S255A by pET25b in Escherichia coli BL21-Codon-Plus(DE3) Thermococcus kodakarensis

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.4.21.B57 crystallization of the active-site mutant S255A of pro-Tk-subtilisin. The crystal is grown at 4°C by the sitting-drop vapour-diffusion method. Native X-ray diffraction data are collected to 2.3 A resolution.They crystal belongs to the orthorhombic space group I222, with unit-cell parameters a = 92.69, b = 121.78, c = 77.53 A. Assuming the presence of one molecule per asymmetric unit, the Matthews coefficient V(M) was calculated to be 2.6 A(3) Da(-1) and the solvent content was 53.1% Thermococcus kodakarensis
3.4.21.62 pro-S255A crystallized in complex with Ca2+, by sitting-drop vapour-diffusion method, to 2.3 A resolution. Crystal belongs to space group I222, with unit cell parameters a = 92.69, b = 121.78, c = 77.53 A. Matthews coefficient is 2.6 A 3 Da-1 and the solvent content is 53.1%. Thermococcus kodakarensis

Protein Variants

EC Number Protein Variants Comment Organism
3.4.21.B57 S255A active-site mutant enzyme Thermococcus kodakarensis
3.4.21.62 S255A active-site mutant of pro-subtilisin. Accumulates in cells in inclusion bodies like pro-subtilisin Thermococcus kodakarensis

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.4.21.62 Ca2+ six calcium ions bind to pro-S255A at the loop regions. The at least six Ca2+ ions bind to pro-S255A too tightly to be removed by dialysis Thermococcus kodakarensis

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.4.21.B57 45000
-
 active-site mutant S255A of pro-Tk-subtilisin Thermococcus kodakarensis
3.4.21.62 45000
-
 pro-S255A, gel filtration Thermococcus kodakarensis

Organism

EC Number Organism UniProt Comment Textmining
3.4.21.B57 Thermococcus kodakarensis P58502 sequence including singnal peptide (amino acid 1-24) and propeptide (amino acid 25-106)
-
3.4.21.62 Thermococcus kodakarensis
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.4.21.B57
-
 Thermococcus kodakarensis
3.4.21.62 pro-S255A purified in the presence of 8 M urea. Refolded by removing urea in the presence of Ca2+. Further purified by gel filtration Thermococcus kodakarensis

Subunits

EC Number Subunits Comment Organism
3.4.21.B57 monomer 1 * 45000, active-site mutant S255A of pro-Tk-subtilisin, SDS-PAGE Thermococcus kodakarensis
3.4.21.62 monomer pro-S255A, SDS–PAGE Thermococcus kodakarensis

Synonyms

EC Number Synonyms Comment Organism
3.4.21.B57 Tk-subtilisin
-
 Thermococcus kodakarensis