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Information on EC 5.4.99.28 - tRNA pseudouridine32 synthase and Organism(s) Escherichia coli and UniProt Accession P0AA37

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EC Tree
     5 Isomerases
         5.4 Intramolecular transferases
             5.4.99 Transferring other groups
                5.4.99.28 tRNA pseudouridine32 synthase
IUBMB Comments
The dual-specificity enzyme from Escherichia coli also catalyses the formation of pseudouridine746 in 23S rRNA . cf. EC 5.4.99.29 (23S rRNA pseudouridine746 synthase).
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This record set is specific for:
Escherichia coli
UNIPROT: P0AA37
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Word Map
The taxonomic range for the selected organisms is: Escherichia coli
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota, Archaea
Reaction Schemes
tRNA uridine32
=
tRNA pseudouridine32
Synonyms
rib2/pus8p, pseudouridine synthase rlua, trna:psi32-synthase, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pseudouridine synthase RluA
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
tRNA uridine32 = tRNA pseudouridine32
show the reaction diagram
SYSTEMATIC NAME
IUBMB Comments
tRNA-uridine32 uracil mutase
The dual-specificity enzyme from Escherichia coli also catalyses the formation of pseudouridine746 in 23S rRNA [5]. cf. EC 5.4.99.29 (23S rRNA pseudouridine746 synthase).
CAS REGISTRY NUMBER
COMMENTARY hide
430429-15-5
-
61506-89-6
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
tRNA uridine32
tRNA pseudouridine32
show the reaction diagram
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
tRNA uridine32
tRNA pseudouridine32
show the reaction diagram
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.000108 - 0.000455
tRNA uridine32
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.051 - 0.15
tRNA uridine32
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
112.1 - 937.5
tRNA uridine32
-
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
the rluA deletion mutant fails to form either 23S RNA pseudouridine746 or tRNA pseudouridine32. Replacement of rluA in trans on a rescue plasmid restores both pseudouridines. There is no difference in exponential growth rate between wild-type and MG1655(rluA-) either in rich or minimal medium at 24, 37, or 42°C, but when both strains are grown together, a strong selection against the deletion strain is observed
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
24432
x * 24432, calculated from sequence
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
x * 24432, calculated from sequence
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
vapor diffusion method. 2.05 A resolution structure of RluA bound to a substrate RNA comprising the anticodon stem loop of tRNAPhe reveals that enzyme binding induces a dramatic reorganization of the RNA
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C117A
kcat is 1.4fold lower than wild-type value, KM is 3.9fold higher than wild-type value
C117A/A128A
kcat is 1.3fold lower than wild-type value, KM is 1.5fold higher than wild-type value
C128A
kcat is 1.5fold higher than wild-type value, KM is 1.5fold higher than wild-type value
P29G
kcat-value is 1.6fold lower than wild-type value. KM-value is 2.9fold higher than wild-type value
P29L
kcat-value is 1.9fold lower than wild-type value. KM-value is 4.2fold higher than wild-type value
additional information
substitution of cysteine for amino acids with nonnucleophilic side chains does not significantly alter the catalytic activity
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
35
Tm-value for mutant enzyme P29L
38
Tm-value for mutant enzyme P29G is 37.9°C
45
Tm-value for wild-type enzyme is 44.9°C
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Ramamurthy, V.; Swann, S.; Spedaliere, C.; Mueller, E.
Role of cysteine residues in pseudouridine synhases of different families
Biochemistry
38
13106-13111
1999
Escherichia coli (P0AA37)
Manually annotated by BRENDA team
Hamilton, C.S.; Greco, T.M.; Vizthum, C.A.; Ginter, J.M.; Johnston, M.V.; Mueller, E.G.
Mechanistic investigations of the pseudouridine synthase RluA using RNA containing 5-fluorouridine
Biochemistry
45
12029-12038
2006
Escherichia coli (P0AA37)
Manually annotated by BRENDA team
Hoang, C.; Chen, J.; Vizthum, C.A.; Kandel, J.M.; Hamilton, C.S.; Mueller, E.G.; Ferre-DAmare, A.R.
Crystal structure of pseudouridine synthase RluA: indirect sequence readout through protein-induced RNA structure
Mol. Cell
24
535-545
2006
Escherichia coli (P0AA37)
Manually annotated by BRENDA team
Spedaliere, C.J.; Hamilton, C.S.; Mueller, E.G.
Functional importance of motif I of pseudouridine synthases: mutagenesis of aligned lysine and proline residues
Biochemistry
39
9459-9465
2000
Escherichia coli (P0AA37)
Manually annotated by BRENDA team
Raychaudhuri, S.; Niu, L.; Conrad, J.; Lane, B.G.; Ofengand, J.
Functional effect of deletion and mutation of the Escherichia coli ribosomal RNA and tRNA pseudouridine synthase RluA
J. Biol. Chem.
274
18880-18886
1999
Escherichia coli (P0AA37), Escherichia coli
Manually annotated by BRENDA team
Wrzesinski, J.; Nurse, K.; Bakin, A.; Lane, B.G.; Ofengand, J.
A dual-specificity pseudouridine synthase: an Escherichia coli synthase purified and cloned on the basis of its specificity for psi 746 in 23S RNA is also specific for psi 32 in tRNA(phe)
RNA
1
437-448
1995
Escherichia coli (P0AA37)
Manually annotated by BRENDA team