A luciferase from dinoflagellates such as Gonyaulax polyedra, Lingulodinium polyedrum, Noctiluca scintillans, and Pyrocystis lunula. It is a single protein with three luciferase domains. The luciferin is strongly bound by a luciferin binding protein above a pH of 7.
A luciferase from dinoflagellates such as Gonyaulax polyedra, Lingulodinium polyedrum, Noctiluca scintillans, and Pyrocystis lunula. It is a single protein with three luciferase domains. The luciferin is strongly bound by a luciferin binding protein above a pH of 7.
each of the three domains of Lingulodinium polyedrum luciferase encodes an active luciferase that catalyzes the oxidation of a chlorophyll-derived open tetrapyrrole (dinoflagellate luciferin) to produce blue light
each of the three domains of Lingulodinium polyedrum luciferase encodes an active luciferase that catalyzes the oxidation of a chlorophyll-derived open tetrapyrrole (dinoflagellate luciferin) to produce blue light
after a 15 min incubation at pH 8.0 and 20°C with subtilisin, trypsin, chymotrypsin, pepsin, carboxypeptidase, and leucine aminopeptidase, the flash height measured in the presence of saturating luciferin is 50% lowered and, after 1 h, the flash height is 10% of the control. Moreover, proteolysis of the extracts completely inactivates the luciferase
after a 15 min incubation at pH 8.0 and 20°C with subtilisin, trypsin, chymotrypsin, pepsin, carboxypeptidase, and leucine aminopeptidase, the flash height measured in the presence of saturating luciferin is 50% lowered and, after 1 h, the flash height is 10% of the control. Moreover, proteolysis of the extracts completely inactivates the luciferase
after a 15 min incubation at pH 8.0 and 20°C with subtilisin, trypsin, chymotrypsin, pepsin, carboxypeptidase, and leucine aminopeptidase, the flash height measured in the presence of saturating luciferin is 50% lowered and, after 1 h, the flash height is 10% of the control. Moreover, proteolysis of the extracts completely inactivates the luciferase
the crude extract has a specific activity of 0.000000258 at pH 8.0 and 20°C and the purified enzyme shows a specific activity of 0.00000348 quanta/min/mg at pH 8.0 and 20°C
the crude extract has a specific activity of 0.000000258 at pH 8.0 and 20°C and the purified enzyme shows a specific activity of 0.00000348 quanta/min/mg at pH 8.0 and 20°C
the crude extract has a specific activity of 0.000000258 at pH 8.0 and 20°C and the purified enzyme shows a specific activity of 0.00000348 quanta/min/mg at pH 8.0 and 20°C
constant pH accelerated molecular dynamics (CpHaMD) is applied to investigate the conformational changes associated with the activation of the enzyme (LCF) upon acidification. The protonation of several residues, including the previously identifies intramolecularly conserved histidines and the H1064/H1065 dyad, correlates with a large scale conformational change in which the N-terminal domain reorganizes to allow the substrate access to the active site
the oxidation of luciferin by Dinoflagellates luciferase only takes place at low pH. Computational tools are used to predict the open structure of the luciferase in Lingulodinium polyedra and to decipher the nature of the opening mechanism. Through accelerated molecular dynamics simulations, it is demonstrated that the closed-open conformational change likely takes place via a tilt of the pH-regulatory helix-loop-helix domain. It is proposed that the molecular basis for the transition is electrostatic repulsion between histidine-cation pairs, which destabilizes the closed conformation at low pH
the oxidation of luciferin by Dinoflagellates luciferase only takes place at low pH. Computational tools are used to predict the open structure of the luciferase in Lingulodinium polyedra and to decipher the nature of the opening mechanism. Through accelerated molecular dynamics simulations, it is demonstrated that the closed-open conformational change likely takes place via a tilt of the pH-regulatory helix-loop-helix domain. It is proposed that the molecular basis for the transition is electrostatic repulsion between histidine-cation pairs, which destabilizes the closed conformation at low pH
in Lingulodinium polyedrum, three components are involved in luminescence: the enzyme luciferase, the substrate luciferin, and a luciferin-binding protein which protects luciferin from autoxidation
domains D2-LCF and D3-LCF, sitting drop vapor diffusion method, using 13% (w/v) PEG 10000, 0.075 M bicine pH 7.7, 25% (v/v) glycerol for domain D2-LCF and 18-20% methyl ether PEG 2000, 100 mM EPPS buffer pH 7.8-8.4 for domain D3-LCF
deletion mutants for N-terminal region of domain 3 of the luciferase show above 20% of wild type activity, the activities of C-terminal deleted mutants decrease drastically to below 1% of wild type
for each luciferase domain (D1, D2, D3) the removal of approximately 50 N-terminal amino acids results in an increase in the ratio of luciferase activity at pH 8.0 relative to that at pH 6.3. At pH 8.0, peptides D1167-486, D2535-897, and D3927-1241, lacking the first 55, 47, and 62 amino acids of their N-termini retain about 40%, 50%, and 70% of the pH 6.3 luciferase activity, respectively
the activity of the full-length native enzyme drops to near zero at pH 8.0, the activity of domain fragments also peaks at pH 6.3 but remains high at 8.0
at 20°C the luciferase is progressively inactivated after a 4 h incubation time at each pH, about 50% residual activity is observed at 30°C, the enzyme appears completely inactive at 40°C and pH 7.9 and shows about 10% residual activity at 40°C and pH 6.7
at 20°C the luciferase is progressively inactivated after a 4 h incubation time at each pH, about 50% residual activity is observed at 30°C, the enzyme appears completely inactive at 40°C and pH 7.9 and shows about 10% residual activity at 40°C and pH 6.7
at 20°C the luciferase is progressively inactivated after a 4 h incubation time at each pH, about 50% residual activity is observed at 30°C, the enzyme appears completely inactive at 40°C and pH 7.9 and shows about 10% residual activity at 40°C and pH 6.7
at 18 and 5°C guanidine-HCl in the concentration range from 2-6 M, denatures irreversibly the luciferase. Luciferin does not protect the luciferase against denaturation by this agent
at 18°C and 5°C guanidine-HCl in the concentration range from 2-6 M, denatures irreversibly the luciferase. Luciferin does not protect the luciferase against denaturation by this agent
at 18 and 5°C guanidine-HCl in the concentration range from 2-6 M, denatures irreversibly the luciferase. Luciferin does not protect the luciferase against denaturation by this agent
at 18 and 5°C urea in the concentration range from 2-8 M, denatures irreversibly the luciferase. Luciferin does not protect the luciferase against denaturation by this agent
at 18°C and 5°C urea in the concentration range from 2-8 M, denatures irreversibly the luciferase. Luciferin does not protect the luciferase against denaturation by this agent
at 18 and 5°C urea in the concentration range from 2-8 M, denatures irreversibly the luciferase. Luciferin does not protect the luciferase against denaturation by this agent
luciferase activity in cell-free extracts of Gonyaulax polyedm undergoes a cyclic daily change such that activities of extracts made in the middle of the night phase may be 10 times greater than in extracts of day phase cells. The circadian rhythm of luciferase activity is a result of biological clock-controlled synthesis and/or degradation of the luciferase polypeptide
luciferase activity in cell-free extracts of Gonyaulax polyedm undergoes a cyclic daily change such that activities of extracts made in the middle of the night phase may be 10 times greater than in extracts of day phase cells. The circadian rhythm of luciferase activity is a result of biological clock-controlled synthesis and/or degradation of the luciferase polypeptide
luciferase activity in cell-free extracts of Gonyaulax polyedm undergoes a cyclic daily change such that activities of extracts made in the middle of the night phase may be 10 times greater than in extracts of day phase cells. The circadian rhythm of luciferase activity is a result of biological clock-controlled synthesis and/or degradation of the luciferase polypeptide