Publications

• Role of a luciferin-binding protein in the circadian bioluminescent reaction of Gonyaulax polyedra.

  Morse D., Pappenheimer A.M. Jr., Hastings J.W.

  A luciferin-binding protein (LBP), which binds and protects from autoxidation the substrate of the circadian bioluminescent reaction of Gonyaulax polyedra, has been purified to near homogeneity. The purified protein is a dimer with two identical 72-kDa subunits, and an isoelectric point of 6.7. LB ... >> More

  A luciferin-binding protein (LBP), which binds and protects from autoxidation the substrate of the circadian bioluminescent reaction of Gonyaulax polyedra, has been purified to near homogeneity. The purified protein is a dimer with two identical 72-kDa subunits, and an isoelectric point of 6.7. LBP is a major component of the cells, comprising about 1% of the total protein during the night phase, but drops to only about 0.1% during the day. The luciferin is protected from autoxidation by binding to LBP, and one luciferin is bound per dimer at alkaline pH (Ka approximately 5 x 10(7) M-1). The protein undergoes a conformational change with release of luciferin at pH values below 7, concurrent with an activation of Gonyaulax luciferase. LBP thus has a dual role in the circadian bioluminescent system. << Less

  J Biol Chem 264:11822-11826(1989) [PubMed] [EuropePMC]

• Crystal structure of a pH-regulated luciferase catalyzing the bioluminescent oxidation of an open tetrapyrrole.

  Schultz L.W., Liu L., Cegielski M., Hastings J.W.

  The luciferase of Lingulodinium polyedrum, a marine bioluminescent dinoflagellate, consists of three similar but not identical domains in a single polypeptide. Each encodes an active luciferase that catalyzes the oxidation of a chlorophyll-derived open tetrapyrrole (dinoflagellate luciferin) to pr ... >> More

  The luciferase of Lingulodinium polyedrum, a marine bioluminescent dinoflagellate, consists of three similar but not identical domains in a single polypeptide. Each encodes an active luciferase that catalyzes the oxidation of a chlorophyll-derived open tetrapyrrole (dinoflagellate luciferin) to produce blue light. These domains share no sequence similarity with any other in the GenBank database and no structural or motif similarity with any other luciferase. We report here the 1.8-A crystal structure of the third domain, D3, at pH 8, and a mechanism for its activity regulation by pH. D3 consists of two major structural elements: a beta-barrel pocket putatively for substrate binding and catalysis and a regulatory three-helix bundle. N-terminal histidine residues previously shown to regulate activity by pH are at the interface of the helices in the bundle. Molecular dynamics calculations indicate that, in response to changes in pH, these histidines could trigger a large molecular motion of the bundle, thereby exposing the active site to the substrate. << Less

  Proc. Natl. Acad. Sci. U.S.A. 102:1378-1383(2005) [PubMed] [EuropePMC]

• The biological clock in Gonyaulax controls luciferase activity by regulating turnover.

  Dunlap J.C., Hastings J.W.

  Luciferase activity in cell-free extracts of the bioluminescent marine dinoflagellate Gonyaulax polyedra 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. These cyclic changes continue ... >> More

  Luciferase activity in cell-free extracts of the bioluminescent marine dinoflagellate Gonyaulax polyedra 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. These cyclic changes continue under constant conditions, in a manner indicative of control by an endogenous circadian cellular mechanism. This paper describes the purification and properties of the higher molecular weight unproteolyzed luciferase from both day and night phase cells. Comparisons of the two preparations with respect to several physicochemical, enzymatic, and immunological criteria were made in order to establish the basis for the activity difference; no differences between day and night species were found. A given amount of antiluciferase inactivated the same amount of luciferase activity in both day and night extracts; their specific activities are therefore the same. These data strongly suggest that the luciferase is the same polypeptide in day and night extracts, and that such extracts contain different amounts of the enzyme. We therefore postulate that the circadian rhythm of luciferase activity is a result of biological clock-controlled synthesis and/or degradation of the luciferase polypeptide. << Less

  J. Biol. Chem. 256:10509-10518(1981) [PubMed] [EuropePMC]

• Dinoflagellate luciferin-binding protein.

  Morse D., Mittag M.

  Methods Enzymol 305:258-276(2000) [PubMed] [EuropePMC]

• Cloning, sequencing and expression of dinoflagellate luciferase DNA from a marine alga, Gonyaulax polyedra.

  Bae Y.M., Hastings J.W.

  The marine dinoflagellate, Gonyaulax polyedra emits light in a reaction involving the enzymatic oxidation of its tetrapyrrole luciferin by molecular oxygen; its luciferase (LCF) single chain has an estimated molecular mass of 130 kDa, and exhibits a circadian rhythm in its activity. A cDNA express ... >> More

  The marine dinoflagellate, Gonyaulax polyedra emits light in a reaction involving the enzymatic oxidation of its tetrapyrrole luciferin by molecular oxygen; its luciferase (LCF) single chain has an estimated molecular mass of 130 kDa, and exhibits a circadian rhythm in its activity. A cDNA expression library in the lambda ZAPII vector was constructed from the polyadenylated RNA isolated from the Gonyaulax cells during the early night phase, the time at which LCF synthesis is believed to be greatest. Of the approx. 1.2 . 10(5) phages from the library screened with antibody against Gonyaulax LCF, 13 positive plaques were obtained. The nucleotide sequences of two of the larger inserts (2.4 kb and 1.6 kb in length), both carrying the poly(A) tail, were determined and found to be identical in the overlapping region. When expressed in Escherichia coli, both cDNA clones produced active luciferase. A Northern hybridization using the cDNA as a probe showed that the length of the lcf mRNA is approx. 4.1 kb, sufficiently long to encode the 130 kDa LCF. Analyses of polymerase chain reaction products, prepared using both the cloned cDNA and Gonyaulax chromosomal DNA as templates, indicated that the cloned region of the luciferase gene does not carry any introns. This represents the first dinoflagellate luciferase to be cloned and sequenced; its deduced amino acid sequence bears no significant homologies with that of any other luciferase, or any other sequence in the data base. << Less

  Biochim. Biophys. Acta 1219:449-456(1994) [PubMed] [EuropePMC]

• Gonyaulax luciferase: gene structure, protein expression, and purification from recombinant sources.

  Li L.

  Methods Enzymol 305:249-258(2000) [PubMed] [EuropePMC]