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Literature summary extracted from

  • Sato, D.; Shiba, T.; Mizuno, S.; Kawamura, A.; Hanada, S.; Yamada, T.; Shinozaki, M.; Yanagitani, M.; Tamura, T.; Inagaki, K.; Harada, S.
    The hyperthermophilic cystathionine gamma-synthase from the aerobic crenarchaeon Sulfolobus tokodaii expression, purification, crystallization and structural insights (2017), Acta Crystallogr. Sect. F, 73, 152-158 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.5.1.48 expressed in Escherichia coli Rosetta-gami (DE3) cells Sulfurisphaera tokodaii
2.5.1.48 expression in Escherichia coli Sulfurisphaera tokodaii

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.5.1.48 hanging drop vapor diffusion method, using either 100 mM acetate pH 3.6, 54% (w/v) 2-methyl-2,4-pentanediol, 200 mM magnesium chloride and 10 mM ammonium sulfate or 100 mM Tris-HCl pH 7.0, 12-18% (w/v) polyethylene glycol 3350, 200 mM sodium citrate and nickel(II) chloride hexahydrate Sulfurisphaera tokodaii
2.5.1.48 three crystal forms from different temperature and pH conditions, collected to 2.2, 2.9 and 2.7 A resolution for forms I, II and II', respectively. Form I crystals (space group P21, unit-cell parameters a = 58.4, b = 149.3, c = 90.2 A, beta = 108.9°) are obtained at 20°C under acidic pH conditions using 2-methyl-2,4-pentanediol. Under basic pH conditions the enzyme crystallizes in form II at 20°C(space group C2221, unit-cell parameters a = 117.7, b = 117.8, c = 251.3 A) and in form II' at 40°C (space group C2221, unit-cell parameters a = 107.5, b = 127.7, c = 251.1 A) using polyethylene glycol 3350 Sulfurisphaera tokodaii

Protein Variants

EC Number Protein Variants Comment Organism
2.5.1.48 F97Y the mutant shows reduced activity compared to the wild type enzyme Sulfurisphaera tokodaii
2.5.1.48 F97Y enzymatic activity at 80°C is similar to wild-type at acidic to neutral pH, and about 50% of wild-type at basic pH Sulfurisphaera tokodaii

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.5.1.48 150000
-
 gel filtration Sulfurisphaera tokodaii

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.5.1.48 O4-succinyl-L-homoserine + L-cysteine Sulfurisphaera tokodaii
-
 L-cystathionine + succinate
-
 ?
2.5.1.48 O4-succinyl-L-homoserine + L-cysteine Sulfurisphaera tokodaii 7
-
 L-cystathionine + succinate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.5.1.48 Sulfurisphaera tokodaii
-
-
-
2.5.1.48 Sulfurisphaera tokodaii 7
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.5.1.48 hydroxyapatite column chromatography and Sephacryl S-300 gel filtration Sulfurisphaera tokodaii
2.5.1.48 recombinant protein, purified to homogeneity by heat treatment followed by hydroxyapatite and gel filtration Sulfurisphaera tokodaii

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.5.1.48 0.4
-
 pH 6.8, 80°C Sulfurisphaera tokodaii
2.5.1.48 1
-
 pH 7.4, 70°C Sulfurisphaera tokodaii
2.5.1.48 1.4
-
 pH 7.5, 80°C Sulfurisphaera tokodaii
2.5.1.48 1.7
-
 pH 9.1, 80°C Sulfurisphaera tokodaii

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.5.1.48 O4-succinyl-L-homoserine + L-cysteine
-
 Sulfurisphaera tokodaii L-cystathionine + succinate
-
 ?
2.5.1.48 O4-succinyl-L-homoserine + L-cysteine
-
 Sulfurisphaera tokodaii 7 L-cystathionine + succinate
-
 ?

Subunits

EC Number Subunits Comment Organism
2.5.1.48 homotetramer 4 * 42000, SDS-PAGE Sulfurisphaera tokodaii
2.5.1.48 tetramer 4 * 42000, SDS-PAGE Sulfurisphaera tokodaii

Synonyms

EC Number Synonyms Comment Organism
2.5.1.48 CGS
-
 Sulfurisphaera tokodaii
2.5.1.48 cystathionine gamma-synthase
-
 Sulfurisphaera tokodaii

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.5.1.48 80
-
 at pH 7.3 Sulfurisphaera tokodaii

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.5.1.48 9.1
-
 at 80°C Sulfurisphaera tokodaii

Cofactor

EC Number Cofactor Comment Organism Structure
2.5.1.48 pyridoxal 5'-phosphate dependent on Sulfurisphaera tokodaii