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Literature summary for 1.13.12.13 extracted from

  • Hall, M.P.; Unch, J.; Binkowski, B.F.; Valley, M.P.; Butler, B.L.; Wood, M.G.; Otto, P.; Zimmerman, K.; Vidugiris, G.; Machleidt, T.; Robers, M.B.; Benink, H.A.; Eggers, C.T.; Slater, M.R.; Meisenheimer, P.L.; Klaubert, D.H.; Fan, F.; Encell, L.P.; Wood, K.V.
    Engineered luciferase reporter from a deep sea shrimp utilizing a novel imidazopyrazinone substrate (2012), Chem. Biol., 7, 1848-1857.
    View publication on PubMedView publication on EuropePMC
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Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli Oplophorus gracilirostris

Protein Variants

Protein Variants Comment Organism
A4E/Q11R/A33K/V44I/A54F/P115E/Q124K/Y138I mutant is 29000fold brighter than mutant N166R. Western blot analysis shows that mutant A4E/Q11R/A33K/V44I/A54F/P115E/Q124K/Y138I is produced more efficiently than mutant N166R in cells. The increased expression is consistent with improved enzyme stability at 37°C, where the half-life of activity retention is increased 65fold over that of mutant N166R Oplophorus gracilirostris
A4E/Q11R/A33K/V44I/A54F/P115E/Q124K/Y138I/Q18L/F54I/F68Y/L72Q/M75K/I90V/L27V/K33N/K43R/Y68D mutant Nluc: Nluc paired with furimazine produces 2.5 millionfold brighter luminescence in mammalian cells relative to Oluc-19 with coelenterazine. The luminescence produced by Nluc decays with a half-life more than 2 h, significantly longer than for mutant A4E/Q11R/A33K/V44I/A54F/P115E/Q124K/Y138I. Nluc increased luminescence is gained mostly through improvements in protein stability, where Nluc shows markedly greater retention of activity in lysates following incubation at 37°C Oplophorus gracilirostris
N166R mutant shows 50% increased stability at 37°C and 3fold higher luminescence intensity Oplophorus gracilirostris

Organism

Organism UniProt Comment Textmining
Oplophorus gracilirostris
-
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
coelenterazine + O2
-
 Oplophorus gracilirostris oxidized coelenterazine + CO2 + hn
-
 ?
furimazine + O2
-
 Oplophorus gracilirostris ?
-
 ?

Synonyms

Synonyms Comment Organism
NanoLuc
-
 Oplophorus gracilirostris
Oluc-19
-
 Oplophorus gracilirostris

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
37
-
 enzyme retains activity in culture medium for more than 15 h at 37°C Oplophorus gracilirostris
55
-
 enzyme shows high physical stability, retaining activity with incubation up to 55°C Oplophorus gracilirostris