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Literature summary for 2.8.1.15 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
Search for more literature for 2.8.1.15

Cloned(Commentary)

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

Crystallization (Commentary)

Crystallization (Comment) Organism
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

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

Metals/Ions

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

Molecular Weight [Da]

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

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
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
-
 ?
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
-
 ?
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
-
 ?
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

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

Purification (Commentary)

Purification (Comment) Organism
-
 Pyrococcus horikoshii

Storage Stability

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

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
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
-
 ?
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
-
 ?
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
-
 ?
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

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

Synonyms

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