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Literature summary extracted from

  • Cacciapuoti, G.; Servillo, L.; Moretti, M.A.; Porcelli, M.
    Conformational changes and stabilization induced by phosphate binding to 5'-methylthioadenosine phosphorylase from the thermophilic archaeon Sulfolobus solfataricus (2001), Extremophiles, 5, 295-302.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.4.2.28 overexpression in Escherichia coli strain RB791, amino acid determination, incorrect positioning of disulfide bonds, the recombinant enzyme is less thermostable and thermophilic than the native enzyme Saccharolobus solfataricus

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.4.2.28 from recombinant enzyme, hanging drop-vapour diffusion method, protein solution, 7-10 mg/ml, 18°C, reservoir solution for native crystals: Tris-HCl 10 mM, pH 7.4, 28-30% dioxane, 12% 2-methyl-2,4-pentanediol, 0.12 M MgCl2, 0,04 M NaCl, for crystals of enzyme complexed with substrates or sulfate and phosphate ions, substrates are added and NaCl is exchanged for MgSO4 or NH4Cl and KH2PO4, respectively, X-ray structure determination and analysis Saccharolobus solfataricus

General Stability

EC Number General Stability Organism
2.4.2.28 phosphate, and less efficiently also arsenate and sulfate, stabilize the recombinant enzyme against inactiviation by temperature, SDS, urea, and proteolytic enzymes Saccharolobus solfataricus
2.4.2.28 recombinant enzyme, 90°C in 2% SDS, 30 min, loss of 60% activity Saccharolobus solfataricus
2.4.2.28 recombinant enzyme, 90°C in 8 M urea, 30 min, loss of 70% activity Saccharolobus solfataricus

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.4.2.28 proteinase K recombinant enzyme, 10% remaining activity after 4 h at 37°C, phosphate protects Saccharolobus solfataricus
2.4.2.28 Subtilisin recombinant enzyme, 24% remaining activity after 4 h at 37°C, phosphate protects Saccharolobus solfataricus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.4.2.28 27000
-
 6 * 27000, recombinant enzyme, crystal structure analysis Saccharolobus solfataricus

Organism

EC Number Organism UniProt Comment Textmining
2.4.2.28 Saccharolobus solfataricus
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.4.2.28 recombinant from Escherichia coli Saccharolobus solfataricus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.4.2.28 5'-methylthioadenosine + phosphate binding of phosphate and 5-methylthioribose 1-phosphate to the enzyme induces a conformational transition that stabilizes the folded structure of the enzyme Saccharolobus solfataricus adenine + 5-methylthio-D-ribose 1-phosphate
-
 ?
2.4.2.28 5'-methylthioadenosine + phosphate 5'-methylthioadenosine and adenine form ternary complexes with the enzyme only in presence of phosphate and methylthioribose 1-phosphate, respectively Saccharolobus solfataricus adenine + 5-methylthio-D-ribose 1-phosphate
-
 ?

Subunits

EC Number Subunits Comment Organism
2.4.2.28 hexamer 6 * 27000, recombinant enzyme, crystal structure analysis Saccharolobus solfataricus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
2.4.2.28 100
-
 recombinant enzyme, 1 h, 95% remainig activity in presence of 100 mM phosphate Saccharolobus solfataricus
2.4.2.28 110
-
 recombinant enzyme, 10 min, 50% remaining activity in absence and 90% remaining activity in presence of 100 mM phosphate Saccharolobus solfataricus
2.4.2.28 120
-
 recombinant enzyme, 10 min, no activity in absence and 50% remaining activity in presence of 100 mM phosphate Saccharolobus solfataricus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.4.2.28 7.4
-
 assay at Saccharolobus solfataricus