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Literature summary for 6.1.1.2 extracted from

  • Sapienza, P.J.; Li, L.; Williams, T.; Lee, A.L.; Carter, C.W.
    An ancestral tryptophanyl-tRNA synthetase precursor achieves high catalytic rate enhancement without ordered ground-state tertiary structures (2016), ACS Chem. Biol., 11, 1661-1668 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 6.1.1.2

Cloned(Commentary)

Cloned (Comment) Organism
recombinant expression of FLAG- and His-tagged enzyme in Escherichia coli strains BL21 and HSM174 in inclusion bodies, HSM174 cells produce about 5fold more enzyme protein Geobacillus stearothermophilus

General Stability

General Stability Organism
TrpRS Urzyme has substantially reduced hydrophobic stabilization with non-cooperative melting and cold-denaturation Geobacillus stearothermophilus

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required Geobacillus stearothermophilus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + L-tryptophan + tRNATrp Geobacillus stearothermophilus
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 AMP + diphosphate + L-tryptophyl-tRNATrp
-
 ?
additional information Geobacillus stearothermophilus an ancestral tryptophanyl-tRNA synthetase precursor achieves high catalytic rate enhancement without ordered ground-state tertiary structures ?
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 ?

Organism

Organism UniProt Comment Textmining
Geobacillus stearothermophilus
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-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant solubilized FLAG- and His-tagged enzyme from Escherichia coli strains BL21 and HSM174 by nickel affinity chromatography, gel filtration, and ultrafiltration Geobacillus stearothermophilus

Renatured (Commentary)

Renatured (Comment) Organism
solubilization and refolding of recombinant FLAG- and His-tagged enzyme from Escherichia coli strains BL21 and HSM174 by homogenization in 100 mM sodium phosphate, and 6 M guanidinium hydrochloride, pH 7.2 before purification Geobacillus stearothermophilus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + L-tryptophan + tRNATrp
-
 Geobacillus stearothermophilus AMP + diphosphate + L-tryptophyl-tRNATrp
-
 ?
additional information an ancestral tryptophanyl-tRNA synthetase precursor achieves high catalytic rate enhancement without ordered ground-state tertiary structures Geobacillus stearothermophilus ?
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 ?

Subunits

Subunits Comment Organism
More 15N tryptophanyl-tRNA synthetase Urzyme structure analysis by heteronuclear single quantum coherence (HSQC) NMR spectroscopy supplemented by circular dichroism, thermal melting, and induced fluorescence of bound dye. Circular dichroism analysis demonstrates reversible folding of the alpha-helix formed by 61 of 130 residues Geobacillus stearothermophilus

Synonyms

Synonyms Comment Organism
TrpRS Urzyme
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 Geobacillus stearothermophilus
Tryptophanyl-tRNA synthetase
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 Geobacillus stearothermophilus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
additional information
-
 TrpRS Urzyme has substantially reduced hydrophobic stabilization with non-cooperative melting and cold-denaturation Geobacillus stearothermophilus
35
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 a broad maximum stability is centered at about 35°C Geobacillus stearothermophilus

Cofactor

Cofactor Comment Organism Structure
ATP
-
 Geobacillus stearothermophilus

General Information

General Information Comment Organism
evolution an ancestral tryptophanyl-tRNA synthetase precursor achieves high catalytic rate enhancement without ordered ground-state tertiary structures. The TrpRS Urzyme catalytic activity arises neither from tiny amounts of wild-type enzyme, nor from a separate population of folded and highly active Urzyme molecules not in equilibrium with the general population. AaRS Urzymes lack much of the mass of modern aaRS, retaining only a small portion of the hydrophobic cores of the full-length enzymes. AaRS Urzymes contain 120-130 amino acids, and consist of little more than is required to form intact active sites. They retain over 60% of the transition-state stabilization free energy for amino acid activation and the ability to aminoacylate tRNA. Further, they preserve about 20% of the Gibbs energies necessary to discriminate between competing amino acid substrates and preferentially activate amino acids from within, rather than outside, their own class. A major fraction of TrpRS Urzyme molecules contribute to the rate acceleration by transiently forming tight transition-state complexes Geobacillus stearothermophilus
additional information 15N tryptophanyl-tRNA synthetase Urzyme structure analysis by heteronuclear single quantum coherence (HSQC) NMR spectroscopy supplemented by circular dichroism, thermal melting, and induced fluorescence of bound dye. TrpRS Urzyme is not a typical protein domain. Transition state stabilization and catalytic activity from molten globules, overview Geobacillus stearothermophilus