1.13.12.24: calcium-regulated photoprotein
This is an abbreviated version!
For detailed information about calcium-regulated photoprotein, go to the full flat file.
Word Map on EC 1.13.12.24
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1.13.12.24
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photoproteins
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bioluminescence
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aequorin
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luminescence
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obelia
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longissima
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coelenteramide
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ctenophore
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coelenterate
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jellyfish
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ef-hand
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2-hydroperoxycoelenterazine
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mnemiopsis
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hydroid
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clytin
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hydromedusan
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leidyi
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synthesis
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analysis
- 1.13.12.24
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photoproteins
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bioluminescence
- aequorin
-
luminescence
- obelia
- longissima
- coelenteramide
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ctenophore
-
coelenterate
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jellyfish
-
ef-hand
- 2-hydroperoxycoelenterazine
- mnemiopsis
- hydroid
- clytin
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hydromedusan
- leidyi
- synthesis
- analysis
Reaction
Synonyms
aequorin, alcium-activated photoprotein, berovin, BFP-aq, blue fluorescent protein, Ca2+-binding photoprotein, Ca2+-regulated photoprotein, clytin, halistaurin, mitrocomin, mnemiopsin, mnemiopsin 1, mnemiopsin 2, mnemiopsin1, obelin, phialidin
ECTree
1.13.12.24 calcium-regulated photoproteinAdvanced search results
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results (Engineering
Engineering on EC 1.13.12.24 - calcium-regulated photoprotein
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PROTEIN VARIANTS 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
C145S/C152S/C180S
21% of wild-type activity. Contrary to wild-type, 49% residual activity in absence of 2-mercaptoethanol. The regeneration of the triple mutant aequorin is sharply inhibited by 2-mercaptoethanol
C145S/C180S
68% of wild-type activity. Contrary to wild-type, 28% residual activity in absence of 2-mercaptoethanol
C152S/C180S
17% of wild-type activity. Contrary to wild-type, 14% residual activity in absence of 2-mercaptoethanol
D119A
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mutant is an active photoprotein, Ca2+ affinity is reduced by a factor of 20 compared to the wild type
D153G
shift of maximum luminescence to shorter wavelengths. t1/2 is decreased with respect to the half-life time of the native protein (0.7 s). Mutation results in a significant decrease in stability at 65°C
F149
residue participates in stabilization of the coelenterazine peroxide and the triggering of photon emission by linking the third EF-hand to Trp-129 and His-169 coelenterazine binding residues
L170I
16fold increase of wild-type activity, an increase of the photoprotein lifetime at 37°C and increased thermostability
Q168
residue participates in stabilization of the coelenterazine peroxide and the triggering of photon emission by linking the third EF-hand to Trp-129 and His-169 coelenterazine binding residues
Q168R
11fold increase of wild-type activity, increase of the photoprotein lifetime at 37°C and increased thermostablility
W86F
shift of maximum luminescence to shorter wavelengths. The single mutant reaches the highest stability against thermal shock
W86F/D153G
decreased t1/2-value. Maximum emission spectrum at 401 nm. Mutation results in a significant decrease in stability at 65°C
Y82F
Y82F/D153G
decreased t1/2-value. Maximum emission spectrum at 478 nm
Y82F/W86F
maximum emission spectrum at 400 nm. Y82F mutation results in shift of emission to longer wavelength, while the W86F mutation shifts the emission to shorter wavelengths. Compared to wild type aequorin, the Y82F/W86F variant displays a 2fold increase of light half-life. Mutation results in a significant decrease in stability at 65°C
E50G
the luminescence activity of the variant is about 17times greater than that of wild-type photoprotein. The activity of E50G variant increases as a result of more flexibility that is brought about by Gly essential for adopting the correct conformation for functional activity. In comparison with wild-type protein, the variant shows higher optimum temperature and calcium sensitivity as well as slower rate of luminesx02cence decay
E50G/D47N
mutation in loop I of mnemiopsin 2 leads to some conformational alterations in the secondary and tertiary structures which affect both the interaction of the photoprotein with the substrate and Ca2+ coordination
E50G/E53T
mutation in loop I of mnemiopsin 2 leads to some conformational alterations in the secondary and tertiary structures which affect both the interaction of the photoprotein with the substrate and Ca2+ coordination
F88H
mutation shifts the bioluminescence maximum from 482 nm for wild-type to 459 nm. 105% of wild-type bioluminescence yield
F88R
mutation shifts the bioluminescence maximum from 482 nm for wild-type to 474 nm. 99% of wild-type bioluminescence yield
F88W
mutation shifts the bioluminescence maximum from 482 nm for wild-type to 477 nm. 100% of wild-type bioluminescence yield
F88Y
mutation shifts the bioluminescence maximum from 482 nm for wild-type to 453 nm. 100% of wild-type bioluminescence yield
additional information
Y82F
mutation leads to removal of a H-bond from Tyr82 to the bound coelenteramide, and shifts its bioluminescence from 469 nm for wild-type to 501 nm. Mutant is stable with good activity and expressible in mammalian cells
Y82F
mutation shifts the peak towards longer wavelengths. Increased half time of initial light by 3.6 s. Mutation results in a significant decrease in stability at 65°C
additional information
mutation of all 3 Cys resiudes to Ser. Cys-free aequorin displays a two-fold lower specific bioluminescence activity but preserves similar activation properties and light emission kinetics compared to the wild-type aequorin. Mutant aequorin shows increased conformational flexibility
additional information
mutations with increased luminescence intensity neighbor the His-16 or His-169 coelenterazine binding residues or are located in the first EF-hand
additional information
replacement of the loop sequence of EF-hand I with other known loop sequences of Ca2+-binding proteins. Mutants replaced with the EF-hand I and EF-hand III from photoproteins show sufficient luminescence activity, but mutants replaced by EF-hands from Renilla luciferin-binding protein and calmodulin do not. The Ca2+-binding affinity to aequorin is reduced by a positive charge at the second position of the canonical EF-hand motif sequence
additional information
the C-terminal proline residue of aequorin is essential for the long-term stability of the bound coelenterazine. Aequorin lacking the proline residue has only 1% of the specific activity of the wild-type after 2 h, and is virtually inactive after 18 h. Replacement of the C-terminal proline residue with histidine or glutamic acid decreases the specific activity to 10 and 190% of that of the wild-type respectively, these variants show half-lives of 2.4 h and 2.3 h, respectively
additional information
mutation of all 5 Cys resiudes to Ser. Cys-free obelin retains only about 10% of the bioluminescence activity of wild-type obelin and binds coelenterazine and forms active photoprotein much less effectively. The mutation drastically changes the bioluminescence kinetics of obelin completely eliminating a fast component from the light signal decay curve. Replacement of Cys residues increases conformational flexibility
