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Literature summary extracted from

  • Nakagawa, H.; Kuratani, M.; Goto-Ito, S.; Ito, T.; Katsura, K.; Terada, T.; Shirouzu, M.; Sekine, S.; Shigi, N.; Yokoyama, S.
    Crystallographic and mutational studies on the tRNA thiouridine synthetase TtuA (2013), Proteins, 81, 1232-1244.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.8.1.15 expressed in Escherichia coli BL21 Thermus thermophilus
2.8.1.15 expressed in Escherichia coli Rosetta DE3 Pyrococcus horikoshii

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.8.1.15 sitting drop vapor diffusion method at 208C, using 1 microl of sample solution mixed with an equal volume of reservoir solution, composed of 100 mM Bicine–NaOH buffer (pH 9.0) containing 200 mM NaCl, 12% (w/v) polyethylene glycol(PEG) 3350, and 0–10% (v/v) N,N-dimethylformamide. The native crystal belongs to the space group P43212, with unit cell parameters of a 5 b 5 70.1 Angstrom and c 5 128.6 Angstrom Pyrococcus horikoshii

Protein Variants

EC Number Protein Variants Comment Organism
2.8.1.15 C130S ca. 10% of wild-type activity Thermus thermophilus
2.8.1.15 C133S ca. 1% of wild-type activity Thermus thermophilus
2.8.1.15 C222S ca. 5-7% of wild-type activity Thermus thermophilus
2.8.1.15 D161A no activity detected Thermus thermophilus
2.8.1.15 D59A no activity detected Thermus thermophilus
2.8.1.15 E203A ca. 20% of wild-type activity Thermus thermophilus
2.8.1.15 H157A ca. 80% of wild-type activity Thermus thermophilus
2.8.1.15 K58A ca. 62% of wild-type activity Thermus thermophilus
2.8.1.15 N158A ca. 75% of wild-type activity Thermus thermophilus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.8.1.15 Zinc each subunit contains tow N- and c-terminal zinc fingers Pyrococcus horikoshii

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.8.1.15 35600
-
-
Pyrococcus horikoshii

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.8.1.15 ATP + [TtuB sulfur-carrier protein]-Gly-NH-CH2-C(O)SH + 5-methyluridine54 in tRNA + acceptor Thermus thermophilus
-
AMP + diphosphate + 5-methyl-2-thiouridine54 in tRNA + [TtuB sulfur-carrier protein] + reduced acceptor
-
?
2.8.1.15 ATP + [TtuB sulfur-carrier protein]-Gly-NH-CH2-C(O)SH + 5-methyluridine54 in tRNA + acceptor Pyrococcus horikoshii
-
AMP + diphosphate + 5-methyl-2-thiouridine54 in tRNA + [TtuB sulfur-carrier protein] + reduced acceptor
-
?
2.8.1.15 ATP + [TtuB sulfur-carrier protein]-Gly-NH-CH2-C(O)SH + 5-methyluridine54 in tRNA + acceptor Pyrococcus horikoshii DSM 12428
-
AMP + diphosphate + 5-methyl-2-thiouridine54 in tRNA + [TtuB sulfur-carrier protein] + reduced acceptor
-
?
2.8.1.15 ATP + [TtuB sulfur-carrier protein]-Gly-NH-CH2-C(O)SH + 5-methyluridine54 in tRNA + acceptor Thermus thermophilus DSM 7039
-
AMP + diphosphate + 5-methyl-2-thiouridine54 in tRNA + [TtuB sulfur-carrier protein] + reduced acceptor
-
?

Organism

EC Number Organism UniProt Comment Textmining
2.8.1.15 Pyrococcus horikoshii O58038
-
-
2.8.1.15 Pyrococcus horikoshii DSM 12428 O58038
-
-
2.8.1.15 Thermus thermophilus Q72LF3
-
-
2.8.1.15 Thermus thermophilus DSM 7039 Q72LF3
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.8.1.15
-
Pyrococcus horikoshii

Storage Stability

EC Number Storage Stability Organism
2.8.1.15 -80°C, pH 7.0, protein concentration 10 mg/ml Pyrococcus horikoshii

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.8.1.15 ATP + [TtuB sulfur-carrier protein]-Gly-NH-CH2-C(O)SH + 5-methyluridine54 in tRNA + acceptor
-
Thermus thermophilus AMP + diphosphate + 5-methyl-2-thiouridine54 in tRNA + [TtuB sulfur-carrier protein] + reduced acceptor
-
?
2.8.1.15 ATP + [TtuB sulfur-carrier protein]-Gly-NH-CH2-C(O)SH + 5-methyluridine54 in tRNA + acceptor
-
Pyrococcus horikoshii AMP + diphosphate + 5-methyl-2-thiouridine54 in tRNA + [TtuB sulfur-carrier protein] + reduced acceptor
-
?
2.8.1.15 ATP + [TtuB sulfur-carrier protein]-Gly-NH-CH2-C(O)SH + 5-methyluridine54 in tRNA + acceptor
-
Pyrococcus horikoshii DSM 12428 AMP + diphosphate + 5-methyl-2-thiouridine54 in tRNA + [TtuB sulfur-carrier protein] + reduced acceptor
-
?
2.8.1.15 ATP + [TtuB sulfur-carrier protein]-Gly-NH-CH2-C(O)SH + 5-methyluridine54 in tRNA + acceptor
-
Thermus thermophilus DSM 7039 AMP + diphosphate + 5-methyl-2-thiouridine54 in tRNA + [TtuB sulfur-carrier protein] + reduced acceptor
-
?

Subunits

EC Number Subunits Comment Organism
2.8.1.15 homodimer 2 * 35366, calculated from aminoa acid sequence Pyrococcus horikoshii

Synonyms

EC Number Synonyms Comment Organism
2.8.1.15 PH0300
-
Pyrococcus horikoshii
2.8.1.15 TtuA
-
Thermus thermophilus
2.8.1.15 TtuA
-
Pyrococcus horikoshii
2.8.1.15 TT_C0106
-
Thermus thermophilus
2.8.1.15 Veg136 protein
-
Thermus thermophilus