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Literature summary for 2.7.7.87 extracted from

  • Perrochia, L.; Crozat, E.; Hecker, A.; Zhang, W.; Bareille, J.; Collinet, B.; van Tilbeurgh, H.; Forterre, P.; Basta, T.
    In vitro biosynthesis of a universal t6A tRNA modification in Archaea and Eukarya (2013), Nucleic Acids Res., 41, 1953-1964.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 2.7.7.87

Cloned(Commentary)

Cloned (Comment) Organism
a polycistronic sequence containing the genes encoding the PaKEOPS protein subunits Kae, Bud32, Cgi121 and Pcc1 synthesized and cloned into pET28(a) plasmid and transformed in Escherichia coli, Pcc1 fused to a hexa-histidine tag, PaSua5 cloned into pET26b Pyrococcus abyssi
ScSua5 gene amplified by PCR using genomic DNA from Saccharomyces cerevisiae, fused to a hexa-histidine tag at its 3'-end and cloned into pET21d vector and transformed in Escherichia coli Saccharomyces cerevisiae

Metals/Ions

Metals/Ions Comment Organism Structure
Mn2+ stimulating effect, it increases the yield of the reaction to 38% Saccharomyces cerevisiae
Mn2+ stimulating effect, it increases the yield of the reaction to 50% Pyrococcus abyssi

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
90500
-
 theoretical mass of PaKEOPS complex Pyrococcus abyssi
200000
-
 recombinant PaKEOPS complex consisting of Kae1, Bud32, Cgi121 and Pcc1, gel filtration Pyrococcus abyssi

Organism

Organism UniProt Comment Textmining
Pyrococcus abyssi
-
-
-
Saccharomyces cerevisiae
-
-
-

Purification (Commentary)

Purification (Comment) Organism
gravity-flow chromatography on an Ni-NTA resin column, not associated with tRNAs when purified Saccharomyces cerevisiae
gravity-flow chromatography on an Ni-NTA resin column, PaKEOPS complex co-purified with nucleic acids Pyrococcus abyssi

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-threonine + ATP + bicarbonate PaSua5 and PaKEOPS together catalyse the biosynthesis of L-threonylcarbamoyladenylate i.e. t6A Pyrococcus abyssi L-threonylcarbamoyladenylate + diphosphate + H2O
-
 ?
L-threonine + ATP + bicarbonate ScSua5 and ScKEOPS together catalyse the biosynthesis of L-threonylcarbamoyladenylate i.e. t6A Saccharomyces cerevisiae L-threonylcarbamoyladenylate + diphosphate + H2O
-
 ?

Subunits

Subunits Comment Organism
dimer 2 * 90500 Pyrococcus abyssi

Synonyms

Synonyms Comment Organism
PaKEOPS protein complex consisting of Kae1, Bud32, Cgi121 and Pcc1 Pyrococcus abyssi
PaSua5
-
 Pyrococcus abyssi
ScKEOPS protein complex Saccharomyces cerevisiae
ScSua5
-
 Saccharomyces cerevisiae

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
-
 Saccharomyces cerevisiae
50
-
-
 Pyrococcus abyssi

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
additional information
-
 65% of activity still retained at 73°C Pyrococcus abyssi

General Information

General Information Comment Organism
physiological function L-threonylcarbamoyladenylate is a modified nucleotide found in all tRNAs decoding codons starting with adenosine, its role is to facilitate codon–anticodon pairing and to prevent frameshifting during protein synthesis Saccharomyces cerevisiae
physiological function L-threonylcarbamoyladenylate is a modified nucleotide found in all tRNAs decoding codons starting with adenosine, its role is to facilitate codon–anticodon pairing and to prevent frameshifting during protein synthesis Pyrococcus abyssi