Literature summary for 5.4.99.25 extracted from

Friedt, J.; Leavens, F.M.; Mercier, E.; Wieden, H.J.; Kothe, U.
An arginine-aspartate network in the active site of bacterial TruB is critical for catalyzing pseudouridine formation (2014), Nucleic Acids Res., 42, 3857-3870.

Search for more literature for 5.4.99.25

Cloned(Commentary)

Cloned (Comment)	Organism
expressed in Escherichia coli as a His-tagged fusion protein	Escherichia coli

Protein Variants

Protein Variants	Comment	Organism
D90A	dissociation constant: 0.4 microM (wild-type: 0.7 microM), rate of pseudouridine formation compared to wild-type: 500fold decreased, rate of tRNA binding (kapp2): 2/sec (wild-type: 4.2/sec)	Escherichia coli
D90E	dissociation constant: 0.4 microM (wild-type: 0.7 microM), rate of pseudouridine formation compared to wild-type: 30fold decreased, rate of tRNA binding (kapp2): 1.5/sec (wild-type: 4.2/sec)	Escherichia coli
D90N	dissociation constant: 0.4 microM (wild-type: 0.7 microM), rate of pseudouridine formation compared to wild-type: 50fold decreased, rate of tRNA binding (kapp2): 1.9/sec (wild-type: 4.2/sec)	Escherichia coli
R181A	dissociation constant: 2 microM (wild-type: 0.7 microM), rate of pseudouridine formation compared to wild-type: more than 20000fold decreased, rate of tRNA binding (kapp2): 4/sec (wild-type: 4.2/sec)	Escherichia coli
R181K	dissociation constant: 0.7 microM (wild-type: 0.7 microM), rate of pseudouridine formation compared to wild-type: 2500fold decreased, rate of tRNA binding (kapp2): 1.9/sec (wild-type: 4.2/sec)	Escherichia coli
R181M	dissociation constant: 0.5 microM (wild-type: 0.7 microM), rate of pseudouridine formation compared to wild-type: more than 20000fold decreased, rate of tRNA binding (kapp2): 3/sec (wild-type: 4.2/sec)	Escherichia coli

Organism

Organism	UniProt	Comment	Textmining
Escherichia coli	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
using Ni-NTA chromatography	Escherichia coli

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
tRNA uridine55	-	Escherichia coli	tRNA pseudouridine55	-	?

Synonyms

Synonyms	Comment	Organism
TruB	-	Escherichia coli

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
20	-	assay at	Escherichia coli

Use of this online version of BRENDA is free under the CC BY 4.0 license. See terms of use for full details.

Release 2023.1 (January 2023)