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H H Seliger

Publications and source records attributed to H H Seliger.

At least 19 recordsLinked to original sources

Complementary DNA coding click beetle luciferases can elicit bioluminescence of different colors.

Eleven complementary DNA (cDNA) clones were generated from messenger RNA isolated from abdominal light organs of the bioluminescent click beetle, Pyrophorus plagiophthalamus. When expressed in Escherichia coli, these clones can elicit bioluminescence that is readily visible. The clones code for luciferases of four types, distinguished by the colors of bioluminescence they catalyze: green (546 nanometers), yellow-green (560 nanometers), yellow (578 nanometers), and orange (593 nanometers). The amino acid sequences of the different luciferases are 95 to 99 percent identical with each other, but are only 48 percent identical with the sequence of firefly luciferase (Photinus pyralis). Because of the different colors, these clones may be useful in experiments in which multiple reporter genes are needed.

Adenosine Triphosphate↗

Bioluminescent click beetles revisited.

In studying beetle bioluminescence in the early 1960s, Dr. McElroy and his colleagues found that the Jamaican click beetle, Pyrophorus plagiophthalamus, was capable of emitting different colours of light. They further found that the luciferin substrate used by this beetle was the same as that in the firefly, demonstrating that the different colours of bioluminescence were due to differences in the structure of the luciferases. We have recently cloned cDNAs from this beetle species which code for at least four different luciferases. The luciferases are distinguishable by their different colours of bioluminescence when expressed in Escherichia coli. The sequence differences between these different luciferases are few, so the amino acids responsible for the different colours of emission must also be few. Through the construction of hybrid luciferases, by rearranging fragments of the original cDNA clones, we have identified some of these amino acid determinants of colour.

Amino Acid Sequence↗

Modification of spectral sensitivities by screening pigments in the compound eyes of twilight-active fireflies (Coleoptera: Lampyridae).

1. ERG S(lambda) were determined in dark-adapted intact preparations of 6 North American firefly species (Photinus collustrans, marginellus, pyralis, macdermotti, scintillans and Bicellonycha wickershamorum) which restrict their flashing activity to twilight hours. The curves possess narrow (1/2 bandwidth = 50-60 nm) peaks in the yellow (560-580 nm) and a shoulder in the violet (370-420 nm), with a marked attenuation (1.4-2.2 log units) of sensitivity in the green (480-530 nm) region of the spectrum (Fig. 1). Two additional species (Photuris potomaca and frontalis) which initiate flashing at twilight and continue on late into the night (twi-night) possess broad sensitivity maxima around 560 nm (Fig. 3). 2. Selective adaptation experiments isolated near-UV and yellow in P. scintillans (Fig. 2). In the dorsal frontal region of the compound eyes in P. frontalis, high sensitivity existed only in the short wavelength region (near-UV and blue) with a maximum in the blue (lambda max 435 nm) (Fig. 4). 3. The in situ MSP absorption spectrum of the screening pigments was determined in preparations of firefly retina. a) Two kinds of dark brown granules were found in the clear zone region. These granules absorb all across the spectrum with a gradual increase in optical density in the shorter wavelength region in P. pyralis (Fig. 5). b) Besides dark granules, pink-to-red colored screening pigments were present in the vicinity of the rhabdoms. The absorption spectra of these pigments determined in five species were narrow (1/2 bandwidth = 50-80 nm) with species-specific differences in their peak absorption in the green at 525 nm, 510 nm, 512 nm and 517 nm in P. scintillans, macdermotti, collustrans and pyralis, respectively (Fig. 6). A similar pigment was found in P. marginellus with a lambda max at 512 nm (Fig. 7). In all cases, transmission increased both at long and short wavelengths, but more sharply in the long wavelength region (Figs. 6 and 7). Hence each twilight-restricted species has its own unique colored screening pigment. A yellow pigment whose absorption spectrum differed from those found in genus Photinus was found in twi-night active Photuris potomaca (lambda max 461 nm) and night-active P. versicolor (lambda max 456 nm). The transmission of the Photuris pigment increased sharply only in the long wave-length region (Fig. 8).(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Efficient singlet oxygen inactivation of firefly luciferase.

Firefly luciferase is inactivated by singlet oxygen at near diffusion controlled rates, 1.9 X 10(9) M-1 s-1, based on direct comparison with the oxidation of L-histidine. The inactivation kinetics are multiphasic. Inactivation is inhibitable by NaN3. Surface-separated-sensitizer (SSS) system in which singlet oxygen is produced above an air gap separating the reaction solution from the Rose Bengal sensitizer, ensuring only Type II reactions, was compared with a Sensitox II system in which the polymer bound Rose Bengal is contained in the reaction solution and both Type I and Type II reactions can occur. A slight stabilization is afforded by MgSO4.

Histidine↗

Microsomal chemiluminescence of benzo[a]pyrene-7,8-dihydrodiol and its synthetic analogues trans- and cis-1-methoxyvinylpyrene.

The cis- and trans-methoxyvinylpyrene (MVP) analogues of benzo[a]pyrene-7,8-dihydrodiol (7,8-diol) produce specific microsomal chemiluminescence comparable to that produced from 7,8-diol in Aroclor-induced rat liver microsome preparations. The chemiluminescence quantum yields, emission spectra, and the concentration and the temporal kinetics of these three substrates have been examined. Radiolabelled 7,8-diol and t-MVP exhibit significant covalent binding (more than 14%) to microsomal protein when metabolized enzymatically. The extreme quenching of the dioxetane chemiluminescence by both microsomes and phosphatidylcholine, as a model phospholipid, implies that despite the low quantum yield (approx. 10(-8) photons per substrate molecule) for microsomal chemiluminescence of these substrates, a significant fraction of their microsomal oxygenations may proceed via a dioxetane pathway.

Animals↗

Induced chemiluminescence of oxidized fatty acids and oils.

The injection of a strong organic base into milligram quantities of fats and oils dissolved in methylene chloride results in a burst of chemiluminescence whose peak intensity is a function of the previous thermal oxidation history and of the degree of unsaturation of the starting material. The flash of this induced chemiluminescence can be 10(8) times higher than the steady-state "spontaneous" chemiluminescence. The kinetics of the induced chemiluminescence are first order in concentration and second order in time. The emission spectrum is broad and extends into the near infrared. A model based on dioxetane chemiluminescence is proposed to explain the observed kinetics.

Fatty Acids↗

A chemiluminescent probe specific for singlet oxygen.

We have synthesized a methoxyvinylpyrene (MVP) in order to model the mechanism for the observed microsomal chemiluminescence of benzo[a]pyrene 7,8-dihydrodiol, the proximate carcinogenic metabolite of benzo[a]pyrene. This MVP analog has been found to be a highly efficient and specific chemiluminescent probe for picomole quantities of singlet oxygen and singlet oxygen equivalents, and it produces significant chemiluminescence when reacted with cytochrome P-450 enzymes.

7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide↗

A chemiluminescence assay specific for the microsomal metabolite, benzo[a]pyrene-7,8-dihydrodiol.

It is possible to assay for trans-7,8-dihydroxy 7,8-dihydrobenzo[a]-pyrene (BP-7,8-dihydrodiol) in complex metabolite mixtures produced during microsomal metabolism of benzo[a]pyrene (BP) because only the BP-7,8-dihydrodiol metabolite will produce significant chemiluminescence (CL) in the NaOCl-H2O2 singlet oxygen-generating system. The limiting CL sensitivity is 30 pmol in a 1-ml CL reaction mixture. CL assays for BP-7,8-dihydrodiol in microsomal reaction solutions gave concentrations identical with those determined by calibrated high-performance liquid chromatography.

Animals↗

The chemical mimicking of the mechanical stimulation, photoinhibition, and recovery from photoinhibition of bioluminescence in marine dinoflagellate, Gonyaulax polyedra.

Mechanically stimulable bioluminescence and photoinhibition of sensitivity to mechanical stimulation in the marine dinoflagellate Gonyaulax polyedra can be mimicked by a number of cations, proportional to the logarithm of their external concentrations. The data are consistent with mechanical stimulability as a membrane depolarization resulting in an increase in H+ ions at bioluminescence sites and with photoinhibition as a hyperpolarization of the cell membrane.

Animals↗

Specificity of chemiluminescence in the metabolism of benzo[a]pyrene to its carcinogenic diol epoxide.

The metabolism of 7,8-dihydroxy-7,8-dihydrobenzo[a]pyrene results primarily in the ultimate carcinogenic metabolite of benzo[a]pyrene, 7,8-dihydroxy-9,10-oxy-7,8,9,10-tetrahydrobenzo[a]pyrene and to a lesser extent 7,8-dihydroxy-9,10-dioxetane-7,8,9,10-tetrahydrobenzo[a]pyrene, from which chemiluminescence is observed. This specific microsomal chemiluminescence has been used to establish that the rate-limiting reaction in the metabolism of benzo[a]pyrene to the bay region diol epoxide is the production of the 7,8-diol. The microsome-mediated chemiluminescence of the parent benzo[a]pyrene is therefore an indicator of the activity of the specific sequence of metabolic reactions leading to the ultimate carcinogenic metabolite.

7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide↗