Information on EC 1.13.12.5 - Renilla-luciferin 2-monooxygenase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
1.13.12.5
-
RECOMMENDED NAME
GeneOntology No.
Renilla-luciferin 2-monooxygenase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
Renilla luciferin + O2 = oxidized Renilla luciferin + CO2 + hnu
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
decarboxylation
Renilla sp.
-
-
oxidation
oxidative decarboxylation
-
-
redox reaction
-
-
-
-
reduction
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
Renilla-luciferin:oxygen 2-oxidoreductase (decarboxylating)
From the soft coral coelenterate Renilla reniformis. The luciferin is bound to a luciferin-binding protein (BP-LH2). The bioluminescent reaction between the luciferin complex, luciferase and oxygen is triggered by calcium ions. In vivo, the excited state luciferin---luciferase complex undergoes the process of nonradiative energy transfer to an accessory protein, Renilla green fluorescent protein, which results in green bioluminescence. In vitro, Renilla luciferase emits blue light in the absence of any green fluorescent protein.
CAS REGISTRY NUMBER
COMMENTARY hide
346421-46-3
Gaussia luciferase
61869-41-8
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
Renilla sp.
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
-
Renilla luciferase is an enzyme that is responsible for the bioluminescence of the soft coral Renilla
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
12-benzyl-8-hydroxy-2-(4-hydroxybenzyl)-5,11-dihydrobenzo[f]imidazo[1,2-a]quinoxalin-3(6H)-one + O2
oxidized 12-benzyl-8-hydroxy-2-(4-hydroxybenzyl)-5,11-dihydrobenzo[f]imidazo[1,2-a]quinoxalin-3(6H)-one + CO2 + hnu
show the reaction diagram
-
-
-
-
?
2,8-dibenzyl-6-(4-hydroxyphenyl)imidazo[1,2-a]pyrazin-3(7H)-one + O2
oxidized 2,8-dibenzyl-6-(4-hydroxyphenyl)imidazo[1,2-a]pyrazin-3(7H)-one + CO2 + hn
show the reaction diagram
-
-
-
-
?
2-hydroperoxycoelenterazine + O2
?
show the reaction diagram
-
-
-
?
3iso-coelenterazine + O2
?
show the reaction diagram
3me-coelenterazine + O2
?
show the reaction diagram
3meo-coelenterazine + O2
?
show the reaction diagram
8-benzyl-2-(4-fluorobenzyl)-6-(4-hydroxyphenyl)imidazo[1,2-a]pyrazin-3(7H)-one + O2
oxidized 8-benzyl-2-(4-fluorobenzyl)-6-(4-hydroxyphenyl)imidazo[1,2-a]pyrazin-3(7H)-one + CO2 + hnu
show the reaction diagram
-
-
-
-
?
8-benzyl-2-(4-hydroxybenzyl)-6-(4-hydroxyphenyl)imidazo[1,2-a]pyrazin-3(7H)-one + O2
oxidized 8-benzyl-2-(4-hydroxybenzyl)-6-(4-hydroxyphenyl)imidazo[1,2-a]pyrazin-3(7H)-one + CO2 + hn
show the reaction diagram
-
-
-
-
?
alphameh-coelenterazine + O2
?
show the reaction diagram
b-coelenterazine + O2
?
show the reaction diagram
Renilla sp.
-
-
-
-
?
beetle D-luciferin
?
show the reaction diagram
-
450 microM
-
-
?
benzylluciferin + O2
oxidized benzylluciferin + CO2 + hv
show the reaction diagram
-
-
-
-
?
benzylluciferin methyl ether + O2
oxidized benzylluciferin methyl ether + CO2 + hv
show the reaction diagram
-
-
-
-
?
bis-coelenterazine + O2
hnu + ?
show the reaction diagram
-
assay at pH 7.6, about 100fold reduced relative luminescence
-
-
?
cf3-coelenterazine + O2
?
show the reaction diagram
coelenterate-type luciferin + O2
oxidized coelenterate-type luciferin + CO2 + hv
show the reaction diagram
-
-
-
-
?
coelenterazine + ?
?
show the reaction diagram
-
20 microM
-
-
?
coelenterazine + O2
?
show the reaction diagram
coelenterazine + O2
coelenteramide + CO2 + hnu
show the reaction diagram
coelenterazine + O2
coelenteramide + CO2 + hv
show the reaction diagram
coelenterazine + O2
oxidized coelenterazine + CO2 + hn
show the reaction diagram
-
-
-
-
?
coelenterazine-H + O2
?
show the reaction diagram
coelenterazine-v + O2
coelenteramide-v + CO2 + hnu
show the reaction diagram
-
increase of substrate coelenterazine stability by ligating it to Ca2+-triggered coelenterazine-binding protein, CBP, from Renilla muelleri, which apparently functions in the organism for stabilizing and protecting coelenterazine from oxidation. The apo-CBP binds coelenterazine-v very rapidly from Ca2+ free solution, similar to that as the native coelenterazine. At low concentrations, coelenterazine-v bound within CBP generates a brighter bioluminescence signal than would free coelenterazine, thereby increasing the assay sensitivity, overview
orange bioluminescence
-
?
coelentrazine + O2
?
show the reaction diagram
-
-
-
-
?
cp-coelenterazine + O2
hnu + ?
show the reaction diagram
-
assay at pH 7.6, about 100fold reduced relative luminescence
-
-
?
D-luciferin + O2
?
show the reaction diagram
D-luciferin + O2 + ATP
oxidized D-luciferin + CO2 + H2O + AMP + diphosphate + hv
show the reaction diagram
e-coelenterazine + O2
hnu + ?
show the reaction diagram
-
assay at pH 7.6, about 100fold reduced relative luminescence
-
-
?
et-coelenterazine + O2
?
show the reaction diagram
f-coelenterazine + O2
hnu + ?
show the reaction diagram
-
assay at pH 7.6, about 100fold reduced relative luminescence
-
-
?
fcp-coelenterazine + O2
hnu + ?
show the reaction diagram
-
assay at pH 7.6, about 100fold reduced relative luminescence
-
-
?
h-coelenterazine + O2
?
show the reaction diagram
h-coelenterazine + O2
hnu + ?
show the reaction diagram
-
assay at pH 7.6m, about 100fold reduced relative luminescence
-
-
?
hcp-coelenterazine + O2
hnu + ?
show the reaction diagram
-
assay at pH 7.6, about 100fold reduced relative luminescence
-
-
?
i-coelenterazine + O2
?
show the reaction diagram
luciferin + O2
?
show the reaction diagram
-
assay at pH 7.4, 25-30C, 10 min
-
-
?
me-coelenterazine + O2
?
show the reaction diagram
meo-coelenterazine + O2
?
show the reaction diagram
MeO-coelenterazine + O2
hnu + ?
show the reaction diagram
-
assay at pH 7.6, about 100fold reduced relative luminescence
-
-
?
n-coelenterazine + O2
hnu + ?
show the reaction diagram
-
assay at pH 7.6, about 100fold reduced relative luminescence
-
-
?
Renilla luciferin + O2
oxidized Renilla luciferin + CO2 + hv
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
Renilla luciferin + O2
oxidized Renilla luciferin + CO2 + hv
show the reaction diagram
-
-
-
-
?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Br-
-
stimulates luminescence activity
Cl-
-
stimulates luminescence activity
Cu2+
-
strong inhibition of luminescence activity
I-
-
stimulates luminescence activity
KCl
-
stimulation, optimum at 0.5 M
Mg2+
-
aequorin binds to Mg2+ under physiological conditions which modulates its light emission. The binding of Mg2+ to aequorin prevents the molecule from aggregating and stabilizes it in the monomeric form. Aequorin has two Mg2+ binding sites located within the EF-hands
NaCl
-
stimulation, optimum at 0.5 M
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(2Z)-3-[(2-bromophenyl)amino]-1-pyridin-2-ylprop-2-en-1-one
-
-
(2Z)-3-[[4-(dimethylamino)cyclohexa-1,5-dien-1-yl]amino]-1-phenylprop-2-en-1-one
-
-
(3Z)-3-[[4-(dimethylamino)phenyl]methylidene]-1,3-dihydro-2H-indol-2-one
-
common name SU4312
2-(4-ethoxyphenyl)-4-[(4-methylpiperazin-1-yl)carbonyl]quinoline
-
-
2-(4-ethoxyphenyl)-4-[(4-pyridin-2-ylpiperazin-1-yl)carbonyl]quinoline
-
-
2-(4-methylphenyl)-4-[(4-pyrimidin-2-ylpiperazin-1-yl)carbonyl]quinoline
-
-
2-(5-naphthalen-2-yl-1,2,4-oxadiazol-3-yl)pyridine
-
-
2-acetylaminofluorene
Renilla sp.
-
-
2-aminoanthracene
Renilla sp.
-
-
2-Benzylamidopyrazine
-
-
2-hydroxy-N'-[(1E)-(2-hydroxyphenyl)methylidene]benzohydrazide
-
common name SCS
2-mercaptoethanol
2-nitrofluorene
Renilla sp.
-
-
2-[5-(3-chlorophenyl)-1,2,4-oxadiazol-3-yl]pyridine
-
-
4-(1,4-dioxa-8-azaspiro[4.5]dec-8-ylcarbonyl)-2-(4-ethoxyphenyl)quinoline
-
-
4-acetylaminofluorene
Renilla sp.
-
-
4-amino-6-[(E)-[4'-[(Z)-(8-amino-1-hydroxy-5,7-disulfonato-6,7-dihydronaphthalen-2-yl)diazenyl]-3-methylbiphenyl-4-yl]diazenyl]-5-hydroxy-2,3-dihydronaphthalene-1,3-disulfonate
-
common name Evans Blue
4-nitroquinoline-N-oxide
Renilla sp.
-
-
4-[1-(1,3-benzothiazol-2-yl)-2-(4-methylpiperazin-1-yl)-2-oxoethyl]phenol
-
-
6-methyl-2-[(Z)-phenyldiazenyl]pyridin-3-ol
-
common name SIB1757
acetaminophen
Renilla sp.
-
-
actinomycin D
Renilla sp.
-
-
Albendazole
Renilla sp.
-
-
amiodarone
Renilla sp.
-
-
ara C
Renilla sp.
-
-
arsenic III oxide
Renilla sp.
-
-
Benzyl luciferyl sulfate
-
-
Benzyl oxyluciferin
-
-
benzylamine
-
-
bleomycin
Renilla sp.
-
-
brefeldinA
-
5 microg/ml, 24 h, 90% decrease of Gluc secretion
carbamzepine
Renilla sp.
-
-
chlorambucil
Renilla sp.
-
-
chlorpromazine
Renilla sp.
-
-
cisplatin
Renilla sp.
-
-
coelenteramide
CuCl2
-
1 microM, inhibition of 95%
cyclophosphamide
Renilla sp.
-
-
cyclosporin A
Renilla sp.
-
-
cytochalasin B
Dibenzyl amine
-
-
diethyldicarbonate
-
digoxin
Renilla sp.
-
-
dimethylnitrosamine
Renilla sp.
-
-
dithiothreitol
ethyl 4-[[2-(4-ethoxyphenyl)quinolin-4-yl]carbonyl]piperazine-1-carboxylate
-
-
ethyl methanesulfonate
Renilla sp.
-
-
Etioluciferin
-
-
etoposide
Renilla sp.
-
-
gemfibrozil
Renilla sp.
-
-
H-89
Renilla sp.
-
10 microM, reduced luminescence to 50% of RLuc and to 54% of RLuc8
haloperidol
Renilla sp.
-
-
Luciferyl sulfate
-
-
methyl oxyluciferin
-
-
methylmethane sulfonate
Renilla sp.
-
-
mitomycin
Renilla sp.
-
-
monensin A
-
3 microg/ml, 24 h, 65% decrease of Gluc secretion
N'-(3-chlorophenyl)-N-[(1Z)-(3-chlorophenyl)methylidene]imidoformamide
-
common name DCB
N-benzyl-4-methylbenzenesulfonamide
-
inhibition of purified enzyme, inducing activity in cell-based assay
N-methyl-N'-nitro-nitroso-guanidin
Renilla sp.
-
-
nefazodone
Renilla sp.
-
-
nocodazole
-
10 microg/ml, 24 h, 75% decrease of Gluc secretion
oxyluciferin
-
competitive
p-Anidisine
-
-
-
p-Benzyloxyaniline
-
-
phenytoin
Renilla sp.
-
-
Sodium azide
Renilla sp.
-
-
tacrine
Renilla sp.
-
-
taxol
Renilla sp.
-
-
thapsigargin
-
0.03 microM, about 30% decrease of Gluc secretion and activity
troglitazone
Renilla sp.
-
-
tyramine
-
-
Valproic acid
Renilla sp.
-
-
vinblastin
Renilla sp.
-
-
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(1,8-bis-maleimidotriethylene glycol)
Renilla sp.
-
effective conjugation of the N-terminal and C-terminal fragments of Rluc, results in enzymatic activity
-
ATP
Renilla sp.
-
50 microM
bovine serum albumin
-
addition to reaction mixture enhances luciferase activity
-
coelenterazine
-
increase of wild-type Gluc activity at higher substrate concentrations
histamine
Renilla sp.
-
10 microM
Triton X-100
-
0.1%, 3.7fold increase of specific activity of wild-tpye Gluc
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0016 - 0.0133
coelenterazine
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.85
benzylluciferin
Renilla reniformis
-
-
-
0.983
benzylluciferin methyl ether
Renilla reniformis
-
-
-
3.9 - 4.9
coelenterazine
1.85
Luciferin
Renilla reniformis
-
-
0.0167
methyl luciferin
Renilla reniformis
-
-
-
additional information
coelenterazine
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.000022
coelenteramide
-
-
0.0012
N-benzyl-4-methylbenzenesulfonamide
-
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.05
(2Z)-3-[(2-bromophenyl)amino]-1-pyridin-2-ylprop-2-en-1-one
Renilla reniformis
-
IC50 above 0.05 mM
0.011
(2Z)-3-[[4-(dimethylamino)cyclohexa-1,5-dien-1-yl]amino]-1-phenylprop-2-en-1-one
Renilla reniformis
-
; pH 7.2, 22C
0.019
(3Z)-3-[[4-(dimethylamino)phenyl]methylidene]-1,3-dihydro-2H-indol-2-one
Renilla reniformis
-
; pH 7.2, 22C
0.025
2-(4-ethoxyphenyl)-4-[(4-methylpiperazin-1-yl)carbonyl]quinoline
Renilla reniformis
-
IC50 above 0.025 mM
0.011
2-(4-ethoxyphenyl)-4-[(4-pyridin-2-ylpiperazin-1-yl)carbonyl]quinoline
Renilla reniformis
-
; pH 7.2, 22C
0.025
2-(4-methylphenyl)-4-[(4-pyrimidin-2-ylpiperazin-1-yl)carbonyl]quinoline
0.02
2-hydroxy-N'-[(1E)-(2-hydroxyphenyl)methylidene]benzohydrazide
Renilla reniformis
-
; pH 7.2, 22C
0.05
2-[5-(3-chlorophenyl)-1,2,4-oxadiazol-3-yl]pyridine
Renilla reniformis
-
IC50 above 0.05 mM
0.011
4-(1,4-dioxa-8-azaspiro[4.5]dec-8-ylcarbonyl)-2-(4-ethoxyphenyl)quinoline
Renilla reniformis
-
; pH 7.2, 22C
0.01
4-amino-6-[(E)-[4'-[(Z)-(8-amino-1-hydroxy-5,7-disulfonato-6,7-dihydronaphthalen-2-yl)diazenyl]-3-methylbiphenyl-4-yl]diazenyl]-5-hydroxy-2,3-dihydronaphthalene-1,3-disulfonate
Renilla reniformis
-
-
0.0225
4-[1-(1,3-benzothiazol-2-yl)-2-(4-methylpiperazin-1-yl)-2-oxoethyl]phenol
Renilla reniformis
-
; pH 7.2, 22C
0.036
6-methyl-2-[(Z)-phenyldiazenyl]pyridin-3-ol
Renilla reniformis
-
-
0.02
ethyl 4-[[2-(4-ethoxyphenyl)quinolin-4-yl]carbonyl]piperazine-1-carboxylate
Renilla reniformis
-
; pH 7.2, 22C
0.021
H-89
Renilla sp.
-
10 microM coelenterazine, measured RLuc8 activity
0.05
N'-(3-chlorophenyl)-N-[(1Z)-(3-chlorophenyl)methylidene]imidoformamide
Renilla reniformis
-
IC50 above 0.05 mM
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20
-
assay at
25
Renilla sp.
-
assay at
30
-
assay at
37
-
assay at
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
fraction in Escherichia coli cells
-
Manually annotated by BRENDA team
additional information
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
10350
-
MALDI-TOF-MS analyses, hGL-98/168
10740
-
MALDI-TOF-MS analyses, hGL-27/97
16000
Renilla sp.
-
value about, SDS-PAGE, C-terminal fragment
19000
-
catalytic domain
20020
-
MALDI-TOF-MS analyses, hGL
21610
-
calculated from sequence
25000
-
SDS-PAGE, pCold-hGL
30000
Renilla sp.
-
value about, SDS-PAGE, N-terminal fragment
33000 - 38000
33800
gel filtration
37600
-
SDS-PAGE
37700
-
calculated from sequence
38040
-
calculated from sequence, 4red-shift luciferase
44000
-
SDS-PAGE
47000
-
Western blot, fusion protein Gluc-YFP
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dimer or trimer
-
2 * or 3 * 12000, SDS-PAGE
monomer
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
additional information
-
substitution of native Gaussia luciferase signal sequence by that from human interleukin-2 or albumin, decrease in luciferase activity
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
microbatch method is used for crystallization. The crystals of Ca2+-loaded apoaequorin are grown from 0.02 M CaCl2, 30% v/v 2-methyl-2,4-pentanediol, and 0.1 M sodium acetate (pH 4.6) during less than 1 week of incubation at 4C. The maximum size of crystals is 0.35 * 0.3 * 0.25 mm
free enzyme and in complex with coelenterazine,and mutant K25A/E277A. Diffraction to 1.4 A. Structures demonstrate a classic alpha/beta-hydrolase fold. The presumptive catalytic triad residues are D120, E144, and H285. Additionally determination of structure of the accessory green fluorescent protein
molecular docking simulations with the coelenterazine substrate. Two triads of residues are critical for catalysis. Putative catalytic triad residues D120, E144, and H285 bear only limited resemblance to those of aequorin. Residues N53, W121, and P220 are also involved in catalysis
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 10
-
-
439334
6.5 - 8.5
-
below 6.5 and above 8.5 less than 20% residual activity
700221
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4 - 50
-
at 50C after 30 min incubation time only 20% residual activity, at 40C after 1 h incubation time complete loss of activity
37
Renilla sp.
-
mutations Tat229C/A-230 and Tat 229-230 retained higher activity than other mutations
39
temperature for 50% inactivation of aequorin bioluminescence by a 30-min heat shock, mutant enzyme F149S
44
temperature for 50% inactivation of aequorin bioluminescence by a 30-min heat shock, wild-type enzyme and mutant enzyme E35G
46
temperature for 50% inactivation of aequorin bioluminescence by a 30-min heat shock, mutant enzyme E128G
48
temperature for 50% inactivation of aequorin bioluminescence by a 30-min heat shock, mutant enzyme L170I
49
temperature for 50% inactivation of aequorin bioluminescence by a 30-min heat shock, mutant enzyme D117G
51
temperature for 50% inactivation of aequorin bioluminescence by a 30-min heat shock, mutant enzyme D153G
52
temperature for 50% inactivation of aequorin bioluminescence by a 30-min heat shock, mutant enzyme Q168AR/L170M, Q168A/L170V and Q168K/L170V
53
temperature for 50% inactivation of aequorin bioluminescence by a 30-min heat shock, mutant enzyme Q168R
57
temperature for 50% inactivation of aequorin bioluminescence by a 30-min heat shock, mutant enzymes Q68R/L170I and Q168R/L179V
95
-
3 min in the presence of DTT and then cooling on ice, over 93% of the luminescence activity of BFP-aq with coelenterazine is recovered
additional information
Renilla sp.
-
at 37C reduced activity compared with remaining activity at 25C
OXIDATION STABILITY
ORGANISM
UNIPROT
LITERATURE
anaerobic luciferin binding promotes a rapid concentration-dependent aggregation of luciferase, which results in irreversible inactivation of the enzyme
-
439337
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
4C or -80C, BFP-aq and gFP-aq do not show significant loss of their fluorescence and luciferase activities after 1 year
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
by affinity chromatography
-
Hi-Trap column
Renilla sp.
-
souble and insoluble fraction by nickel-chelate column
-
using Ni-chelate column
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
catalytic domain (19 kDa) is expressed using a cold-induced expression system in Escherichia coli as His-tagged fusion proteins
-
expressed in Escherichia coli
-
expressed in HEK-293T cells; expression of RLuc8 in Escherichia coli BL21
Renilla sp.
-
expressed using a cold-induced expression system in Escherichia coli as His-tagged fusion proteins
expression in 293T HEK fibroblasts and HF8 human fibroblasts
-
expression in Arabidopsis; expression in Escherichia coli strain BL21
-
expression in Chlamydomonas reinhardtii
expression in CHO cells and HepG2 cells
-
expression in CHO-K1 cells and PC12D cells
-
expression in Escherichia coli
expression in Escherichia coli BL21
-
expression in Escherichia coli DH10B cells and 293T human kidney fibroblasts
-
expression in Escherichia coli DH5alpha
-
expression in Escherichia coli strain DH5alpha and MCF-7 cells
Renilla sp.
-
expression in Escherichia coli strain ER 2566
Renilla sp.
-
expression in HEK-293 cells or Grip-Tite 293 MSR cells
-
expression in HeLa-cell
-
expression in Hep-G2 cell
Renilla sp.
-
expression in LNCaP cell, pRL-SV40 cell, pRL-null cell
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expression in mouse and human C57/BL6 primary T cells, expression in PG-13 cells
expression in rabbit leukocyte lysate
Renilla sp.
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expression of His-tagged apoclytin into the periplasmic space in Escherichia coli
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expression of Renilla luciferase split fragments in HeLa cells, mitochondrial localization, overview
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expression of stabilized variant RLuc8 with noncleavable N-terminal His6-tag
fragments of Renilla luciferase (Rluc) are fused to the chemotaxis-associated response regulator CheY3 and its phosphatase CheZ in the enteric pathogen Vibrio cholerae in order to demonstrate dynamic protein-protein interactions by luciferase complementation
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D117G
the temperature for 50% inactivation of aequorin bioluminescence by a 30-min heat shock is 5C higher than wild-type value
D153G
the temperature for 50% inactivation of aequorin bioluminescence by a 30-min heat shock is 7C higher than wild-type value
E128G
the temperature for 50% inactivation of aequorin bioluminescence by a 30-min heat shock is 2C higher than wild-type value
E35G
the temperature for 50% inactivation of aequorin bioluminescence by a 30-min heat shock is identical to wild-type value
F149S
the temperature for 50% inactivation of aequorin bioluminescence by a 30-min heat shock is 5C lower than wild-type value
L170I
the temperature for 50% inactivation of aequorin bioluminescence by a 30-min heat shock is 4C higher than wild-type value
Q168A/L170V
the temperature for 50% inactivation of aequorin bioluminescence by a 30-min heat shock is 8C higher than wild-type value
Q168K/L170V
the temperature for 50% inactivation of aequorin bioluminescence by a 30-min heat shock is 8C higher than wild-type value
Q168R
the temperature for 50% inactivation of aequorin bioluminescence by a 30-min heat shock is 9C higher than wild-type value
Q168R/L170I
the temperature for 50% inactivation of aequorin bioluminescence by a 30-min heat shock is 13C higher than wild-type value
Q168R/L170M
the temperature for 50% inactivation of aequorin bioluminescence by a 30-min heat shock is 8C higher than wild-type value
Q168R/L170V
the temperature for 50% inactivation of aequorin bioluminescence by a 30-min heat shock is 13C higher than wild-type value
V44A
the temperature for 50% inactivation of aequorin bioluminescence by a 30-min heat shock is 1C higher than wild-type value
DBHsp-GLuc
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dopamine beta-hydroxlase fused to GLase, lower luminescence activity than wild-type GLuc
deltaSP-GLuc
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GLuc without signal peptide sequence, no significant luminescence
extGLuc
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membrane-anchored
Gluc(HPG)mutant
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contains L-homopropargylglycine, prolonged luminescence, reduced specific activity
M43I
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after 120 s of incubation time still 87% residual light emission compared to only 30% of wild-type Gluc, stabilized light emission with detergent Triton X-100, more stable light emission compared to wild-type Gluc when expression in mammalian cells
pCold-hGL
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two repeat sequences, two catalytic domains
pCold-hGL-27/97
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one repeat sequence, with only one catalytic domain
pCold-hGL-98/168
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one repeat sequence, with only one catalytic domain
pCold-hSGL
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with a signal peptide sequence for secretion
A55T/C124A/C130A/A143M/M253L/S287L/A123T/D154M/E155G/D162E/I163L/V185L
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sequentially introduced into the RM-Luc coding sequence using designed oligonucleotide primers and quick-change site-directed mutagenesis, the mutant RM-Y has a red-shifted bioluminescence spectrum
A164W
73% of wild-type activity
A55T/C124A/S130A/K136R/A143M/M185V/M253L/S287L
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selected mutations enable the protein to emit stronger bioluminescence activity and to be more stable in serum media. Mutant m-Rluc8 exhibits an enhancement in protein expression and shows a 5.6fold improvement in light output, with increased stability in serum media confirmed to last for over 5 days
D120E
1.1% of wild-type activity
D120F
no activity detected
D120Y
no activity detected
E144D
5.6% of wild-type activity
E144F
no activity detected
E144Y
no activity detected
E160N
27.2% of wild-type activity
F180C
14.3% of wild-type activity
F180T
5.4% of wild-type activity
F180Y
11.0% of wild-type activity; 61.8% of wild-type activity
F261A
no activity detected
F261S
no activity detected
H285A
11.3% of wild-type activity
H285D
no activity detected
H285K
no activity detected
H285N
0.1% of wild-type activity
I140L
113% of wild-type activity
I163F
11.0% of wild-type activity
I223W
0.2% of wild-type activity
K189D
24.7% of wild-type activity
K189V
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increased activity
K193S
54.8% of wild-type activity
K25A/E277A
surface mutations made with the intention that they would aid in crystallization, not involved in contacts between proteins in the crystal
K308I
47.5% of wild-type activity
M185V
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increased activity
N53C
3.4% of wild-type activity
N53G
0.5% of wild-type activity
N53H
2.1% of wild-type activity
N53M
1.8% of wild-type activity
N53P
no activity detected
N53Q
25.1% of wild-type activity
N53R
90% of wild-type activity
N53S
20.7% of wild-type activity
P157R
101% of wild-type activity
P220C
72.7% of wild-type activity
P220E
4.9% of wild-type activity
P220F
15.7% of wild-type activity
P220M
140% of wild-type activity
P220Q
222% of wild-type activity
P220S
55.4% of wild-type activity
P220T
89.6% of wild-type activity
P220V
70.5% of wild-type activity
T184C
62.7% of wild-type activity
T184F
46.1% of wild-type activity
T329G
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no significant influence on enzyme activity
V267I
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increased activity
W121A
26.8% of wild-type activity
W121G
4.9% of wild-type activity
W121R
1.1% of wild-type activity
W121S
17.3% of wild-type activity
W121Y
3.1% of wild-type activity
223-224insRev
Renilla sp.
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Rev peptide-inserted Rluc variant
223-224insTat
Renilla sp.
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Tat peptide-inserted Rluc variant
223C/A-224insRev
Renilla sp.
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Tat peptide-inserted Rluc variant
223C/A-224insTat
Renilla sp.
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Tat peptide-inserted Rluc variant
229-230insRev
Renilla sp.
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Rev peptide-inserted Rluc variant
229-230insTat
Renilla sp.
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high thermal stability
229C/A-230insRev
Renilla sp.
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Rev peptide-inserted Rluc variant
229C/A-230insTat
Renilla sp.
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high thermal stability, higher enzyme activity than PI-RLuc
91-92C/AinsRev
Renilla sp.
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higher enzyme activity than PI-RLuc
91-92insRev
Renilla sp.
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Rev peptide-inserted Rluc variant
91-92insTat
Renilla sp.
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higher enzyme activity than PI-RLuc
A55T/C124A/S130A/K136R/A143M/M185V/M253L/S287L
Renilla sp.
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mutant enzyme exhibits a greater than 4-fold enhancement in activity, a 200fold increased resistance to serum inactivation, and a small but measurable 5 nm red shift in the emission spectrum. The enhancement in light output arises from a combination of increases in quantum yield and improvedkinetics
C124A
Renilla sp.
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increased enzymatic activity, 41fold higher luminescence than wild-type luciferase
C124Ac
Renilla sp.
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C-terminal half, decreased enzymatic activity
C124An
Renilla sp.
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N-terminal half, decreased enzymatic activity
C126A
Renilla sp.
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increased activity
extFLuc
Renilla sp.
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membrane-anchored
G229stopstop
Renilla sp.
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Rluc N-terminal fragment
M185V/Q235A
Renilla sp.
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compared with the native enzyme the mutant has twice the rate of inactivation, as measured in murine serum, while incorporating a close to 5fold improvement in light output
N64C
Renilla sp.
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Ca2+-induced interaction between calmodulin and M13 leads to intermolecular complementation of split Renilla luciferase, decreased enzymatic activity
RL8
Renilla sp.
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mutant of Rluc
RLuc8
Renilla sp.
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eight-point mutation variant of renilla luciferase
stop230
Renilla sp.
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Rluc C-terminal fragment
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
biotechnology
molecular biology
pharmacology
additional information