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Literature summary extracted from

  • Ritschel, T.; Atmanene, C.; Reuter, K.; Van Dorsselaer, A.; Sanglier-Cianferani, S.; Klebe, G.
    An integrative approach combining noncovalent mass spectrometry, enzyme kinetics and X-ray crystallography to decipher Tgt protein-protein and protein-RNA interaction (2009), J. Mol. Biol., 393, 833-847.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.4.2.29 mutants introduced into plasmid pET9d-ZM4 and transformed into Escherichia coli BL21(DE3) GOLD cells Zymomonas mobilis

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.4.2.29 mutant K52M, by hanging-drop, vapor-diffusion method and followed by macroseeding, at a resolution of 2.0 A, forms crystals under the same conditions as wild-type, while all attempts to obtain diffracting crystals from mutant Y330F are unsuccessful. Compared to wild-type, crystals of mutant K52M are very fragile and show only a limited diffraction quality Zymomonas mobilis

Protein Variants

EC Number Protein Variants Comment Organism
2.4.2.29 K52M reduced turnover value. At a concentration of protein of 0.01 mM appears almost exclusively as a homodimer as the wild-type, when the concentration of protein is lowered to a minimal value of 0.001 mM, a substantial proportion of monomer becomes evident Zymomonas mobilis
2.4.2.29 Y330F reduced turnover value. At a concentration of protein of 0.01 mM reveals a significant amount of monomer, when the concentration of protein is lowered to a minimal value of 0.001 mM, a substantial proportion of monomer becomes evident Zymomonas mobilis

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.4.2.29 6-amino-2-[(3-piperidin-1-ylpropyl)amino]-1,7-dihydro-8H-imidazo[4,5-g]quinazolin-8-one cannot modify the structure of the dimer interface, prevents the formation of the catalytically active Tgt:tRNA complex, and can disrupt the preformed complex Zymomonas mobilis
2.4.2.29 6-amino-4-[2-(4-methoxyphenyl)ethyl]-1,7-dihydro-8H-imidazo[4,5-g]quinazolin-8-one can modify the structure of the dimer interface, prevents the formation of the catalytically active Tgt:tRNA complex, and can disrupt the preformed complex Zymomonas mobilis
2.4.2.29 6-amino-4-[2-(benzylamino)ethyl]-2-(methylamino)-1,7-dihydro-8H-imidazo[4,5-g]quinazolin-8-one addresses a hydrophobic subpocket close to the dimer interface that may affect quarternary structure formation, prevents the formation of the catalytically active Tgt:tRNA complex, and can disrupt the preformed complex Zymomonas mobilis
2.4.2.29 6-amino-4-[2-[(cyclohexylmethyl)amino]ethyl]-2-(methylamino)-1,7-dihydro-8H-imidazo[4,5-g]quinazolin-8-one can modify the structure of the dimer interface, prevents the formation of the catalytically active Tgt:tRNA complex, and can disrupt the preformed complex Zymomonas mobilis
2.4.2.29 6-amino-4-[2-[(cyclopentylmethyl)amino]ethyl]-2-(methylamino)-1,7-dihydro-8H-imidazo[4,5-g]quinazolin-8-one addresses a hydrophobic subpocket close to the dimer interface that may affect quarternary structure formation, prevents the formation of the catalytically active Tgt:tRNA complex, and can disrupt the preformed complex Zymomonas mobilis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.4.2.29 0.00098
-
[tRNATyr]7-aminomethyl-7-carbaguanine mutant K52M Zymomonas mobilis
2.4.2.29 0.00158
-
[tRNATyr]7-aminomethyl-7-carbaguanine mutant Y330F Zymomonas mobilis
2.4.2.29 0.00217
-
[tRNATyr]7-aminomethyl-7-carbaguanine wild-type Zymomonas mobilis

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.4.2.29 Zn2+ one structural Zn2+ per subunit Zymomonas mobilis

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.4.2.29 85600
-
expected for a Tgt dimer containing one structural Zn2+ per subunit Zymomonas mobilis
2.4.2.29 85610
-
Tgt, noncovalent mass spectrometry Zymomonas mobilis
2.4.2.29 113100
-
Tgt:tRNA complex, noncovalent mass spectrometry Zymomonas mobilis

Organism

EC Number Organism UniProt Comment Textmining
2.4.2.29 Zymomonas mobilis P28720
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.4.2.29 [tRNATyr]7-aminomethyl-7-carbaguanine + guanine Tgt dimer binds specifically to a single tRNA molecule Zymomonas mobilis [tRNATyr]-guanine + 7-aminomethyl-7-carbaguanine
-
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Subunits

EC Number Subunits Comment Organism
2.4.2.29 homodimer noncovalent mass spectrometry, Tgt in solution Zymomonas mobilis

Synonyms

EC Number Synonyms Comment Organism
2.4.2.29 TGT
-
Zymomonas mobilis
2.4.2.29 tRNA-guanine transglycosylase
-
Zymomonas mobilis

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.4.2.29 0.00023
-
[tRNATyr]7-aminomethyl-7-carbaguanine mutant K52M Zymomonas mobilis
2.4.2.29 0.00095
-
[tRNATyr]7-aminomethyl-7-carbaguanine mutant Y330F Zymomonas mobilis
2.4.2.29 0.011
-
[tRNATyr]7-aminomethyl-7-carbaguanine wild-type Zymomonas mobilis