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  • Chemistry  (427)
  • Bacteriorhodopsin  (9)
  • Atomic, Molecular and Optical Physics  (6)
  • 1
    Electronic Resource
    Electronic Resource
    Weinheim : Wiley-Blackwell
    Berichte der deutschen chemischen Gesellschaft 81 (1948), S. 340-353 
    ISSN: 0009-2940
    Keywords: Chemistry ; Inorganic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Es wurde gefunden, daß die kautschukführenden Wurzeln von Taraxacum Kok-saghyz Rod. tyrosinasefreisind, während der mit Koksaghyz nahe verwandte kautschukfreie Löwenzahn (Taraxacum officinale) Tyrosinase enthält. Um festzustellen, welche Beziehung zwischen Kautschuk und Tyrosinase besteht, wurden Hybriden aus beiden Rassen herangezogen, für deren Untersuchung ein colorimetrisches Verfahren angegeben wird, das auch sehr geringe Gehalte von Tyrosinase mit genügender Genauigkeit zu ermitteln gestattet.Das Verfahren beruht auf einer kurzzeitigen Verfolgung der Bildung des roten Farbstoffs aus Tyrosin durch Messung der Lichtabsorption mit dem Photozellencolorimeter nach R. Havemann, während der die in Skalenteilen des Geräts über der Zeit aufgetragenen Reaktionskurven einen geradlinigen Verlauf zeigen. Der Neigungswinkel der Geraden gegen die Abszisse bzw. seine Tangente ist in einem genügend großen Meßbereich der Fermentkonzentration annähernd proportional und kann als relatives Maß für die Aktivität der Fermentpräparate gelten. Eine durch Titration der Reaktionslösungen ermittelte Bezugskurve zwischen dem tg-Wert und dem Tyrosinase-Umsatz nach 2 Stunden führt zu der scheinbaren Wirkungseinheit des Ferments, als die der Umsatz von 1 mg Tyrosin gewählt wird, Woraus sich als absoluter Tyrosinasewirkungswert die Zahl der Wirkungseinheiten/1 g Wurzeltrockensubstanz ergibt.Aus dem Vergleich der Tyrosinasewirkungswerte mit den Kautschukgehalten der Hybriden ergibt sich, daß eine Beziehung zwischen Tyrosinasegehalt und Kautschukgehalt in den Wurzeln nicht besteht. Dadurch ist die Annahme nahegelegt, daß Kautschuk-An-und Abwesenheit sowie Tyrosinase-An- und Abwesenheit für die beiden Stammarten je ein gegensätzliches Merkmalspaar darstellen, die  -  jeweils das eine unabhängig vom anderen - zur Bildung von dihybriden Kreuzungen führen.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Weinheim : Wiley-Blackwell
    Berichte der deutschen chemischen Gesellschaft 81 (1948), S. 354-355 
    ISSN: 0009-2940
    Keywords: Chemistry ; Inorganic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Die Eigenschaft der Tyrosinase, sich in wäßrigen Lösungen bevorzugt im Schaum zu verteilen, wird benutzt, dieses Ferment mit Hilfe einer von Wo. Ostwald beschriebenen kontinuierlich arbeitenden Zerschäumungsvorrichtung anzureichern.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Amsterdam : Elsevier
    FEBS Letters 222 (1987), S. 275-278 
    ISSN: 0014-5793
    Keywords: Bacteriorhodopsin ; EXAFS ; M-intermediate ; Metal-substituted bacteriorhodopsin ; Photocycle
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Biology , Chemistry and Pharmacology , Physics
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 0014-5793
    Keywords: Bacteriorhodopsin ; Membrane protein ; NMR ; Photo-CIDNP ; Proton nuclear resonance ; Side-chain mobility
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Biology , Chemistry and Pharmacology , Physics
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    European biophysics journal 15 (1987), S. 231-236 
    ISSN: 1432-1017
    Keywords: Bacteriorhodopsin ; blue membrane ; purple membrane ; electric dichroism ; cation binding
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Physics
    Notes: Abstract Blue bacteriorhodopsin was prepared by electrodialysis, cation-exchange chromatography and acidification. The electrooptical properties of these preparations compared to those of the native purple bacteriorhodopsin suggest that the blue bacteriorhodopsin has a smaller induced dipole moment than the native purple bacteriorhodopsin and that bound cations in the native bacteriorhodopsin stabilize the protein conformation in the membrane. Purple bacteriorhodopsin was regenerated by addition of potassium, magnesium or ferric ions to blue bacteriorhodopsin. Both spectrscopically and electrooptically the potassium- and ferric-regenerated samples are different from the native purple state. Although the magnesium-regenerated sample is spectroscopically similar to the native purple bacteriorhodopsin, the electrooptical properties are rather similar to those of the cation-depleted blue sample, suggesting that it is very difficult to re-stabilize protein structures once cations are depleted.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1432-1017
    Keywords: Bacteriorhodopsin ; Solid state NMR ; Stable isotope labeling ; Perturbation free analysis of mutants ; Assignment of specific aspartic acids
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Physics
    Notes: Abstract Three mutant strains of Halobacterium sp. GRB with the site of mutation in the bacterioopsin gene (PM 326: Asp96 → Asn; PM 374: Asp96 → Gly; PM 384: Asp85 → Glu) were grown in a synthetic medium containing (4-13C)-Asp. The mutant bacteriorhodopsins labeled with (4-13C)-Asp (37%–45%), and owing to the metabolism of Halobacteria also with (11-13C)-Trp (50%–100%), were isolated as purple membranes and 13C Solid State Magic Angle Sample Spinning (MASS) Nuclear Magnetic Resonance (NMR) spectra of the samples were taken. The Asp96 mutants lacked the signal at 171.3 ppm which was previously assigned to a protonated internal Asp (Engelhard et al. 1989 a). This observation supports the conclusion that Asp96 is protonated in the ground state. PM 384 (Asp85 → Glu) has an absorption maximum at 610 run. It can be converted into a purple form (λmax = 5.40 nm) by treatment with a detergent (CHAPSO). The NMR-spectra of these two species differ from each other and from the wild type. The intensity of the resonance at 173 ppm in the wild type spectrum is reduced in both forms of the mutant protein. It is probable that this signal is caused by Asp85. The amino acid changes result not only in a perturbation of their direct environment but also effects on Trp residues and the chromophore protein interaction can be observed.
    Type of Medium: Electronic Resource
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    European biophysics journal 13 (1986), S. 273-280 
    ISSN: 1432-1017
    Keywords: Bacteriorhodopsin ; blue membrane ; purple membrane films ; electric field-induced states ; electric dichroism
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Physics
    Notes: Abstract Electric field-induced absorption changes of bacteriorhodopsin were studied with different samples of purple membranes which were prepared as randomly oriented and electrically oriented films of purple as well as cation-depleted blue bacteriorhodopsin. The absorption changes were proportional to the square of the field strength up to ≈300 kV/cm. The electric field from the intracellular side to the extracellular side of the purple bacteriorhodopsin induces a spectrum change, resulting in a spectrum similar to that of the cation-depleted blue bacteriorhodopsin. When the field was removed, the purple state was regenerated. The blue state was mainly affected by an electric field in the opposite direction, suggesting a reversible interaction with the Schiff's base bond of the retinal. Since the field-induced reaction of bacteriorhodopsin was observed in the presence of a concomitant steady ion flux, it is assumed that the generation of a local diffusion potential may play an important role in these spectral reactions. Although the fragments were fixed in the dried film, electric dichroism was observed. The dichroic contribution of the total absorbance change was about 15%. The angular displacement of the retinal transition moment was calculated to be 1.5° toward the membrane normal.
    Type of Medium: Electronic Resource
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    European biophysics journal 15 (1988), S. 329-337 
    ISSN: 1432-1017
    Keywords: Bacteriorhodopsin ; blue membrane ; purple membrane films ; electric-field-induced states ; dielectric dispersion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Physics
    Notes: Abstract Electric field induced conformational changes of bacteriorhodopsin were studied in six types of dried film (randomly and electrically oriented membranes of purple as well as cation-depleted blue bacteriorhodopsin) by measuring the frequency dependence of the optical absorbance change and the dielectric dispersion and absorption. For the purple bacteriorhodopsin the optical absorbance change induced by alternating rectangular electric fields of ±300 kV/cm altered the sign twice in the frequency range from 0.001 Hz to 100 kHz (around 0.03 Hz and 100 kHz), indicating that the electric field induced conformational change in these samples consists of, at least, three steps. Similarly, it was found for the blue bacteriorhodopsin that at least two steps are involved. In accord with optical measurements, the dielectric behaviour due to alternating sinusoidal electric fields of±6kV/cm in the frequency range from 10 Hz to 10 MHz showed two broad dispersion/absorption regions, one below 1 kHz and the other around 10–100 kHz. This suggests that the conformational change of bacteriorhodopsin is also reflected by its dielectrical properties and that it is partially induced at 6 kV/cm. Including previous results obtained by analysis of the action of DC fields on purple membrane films, a model for a field-induced cyclic reaction for purple as well as blue bacteriorhodopsin is proposed. In addition it was found that there are electrical interactions among purple membrane fragments in dried films.
    Type of Medium: Electronic Resource
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  • 9
    Electronic Resource
    Electronic Resource
    Springer
    European biophysics journal 18 (1990), S. 63-69 
    ISSN: 1432-1017
    Keywords: Bacteriorhodopsin ; Blue membrane ; Electric-field-induced pH changes ; Proton pump ; Photocycle
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Physics
    Notes: Abstract Electric field induced pH changes of purple membrane suspensions were investigated in the pH range from 4.1 to 7.6 by measuring the absorbance change of pH indicators. In connection with the photocycle and proton pump ability, three different states of bacteriorhodopsin were used: (1) the native purple bacteriorhodopsin (magnesium and calcium ions are bound, the M intermediate exists in the photocycle and protons are pumped), (2) the cation-depleted blue bacteriorhodopsin (no M intermediate), and (3) the regenerated purple bacteriorhodopsin which is produced either by raising the pH or by adding magnesium ions (the M intermediate exists). In the native purple bacteriorhodopsin there are, at least, two types of proton binding sites: one releases protons and the other takes up protons in the presence of the electric field. On the other hand, blue bacteriorhodopsin and the regenerated purple bacteriorhodopsin (pH increase) show neither proton release nor proton uptake. When magnesium ions are added to the suspensions; the field-induced pH change is observed again. Thus, the stability of proton binding depends strongly on the state of bacteriorhodopsin and differences in proton binding are likely to be related to differences in proton pump activity. Furthermore, it is suggested that the appearance of the M intermediate and proton pumping are not necessarily related.
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  • 10
    ISSN: 1432-1017
    Keywords: Bacteriorhodopsin ; Mössbauer spectroscopy ; Cation binding site
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Physics
    Notes: Abstract Bacteriorhodopsin (bR), converted by deionization to the blue form was reconstituted to the active purple membrane by the addition of Fe2+ or Fe3+ ions. 57Fe Mossbauer spectra of these samples were measured at different pH values (pH 3.9, pH 5.0 and pH 7.0) and at temperatures ranging from 4 K to 300 K. The hyperfine parameters reveal two iron environments with oxygen atoms in the neighbourhood of iron. Iron type 1 is in the 3+ high spin state. It is bound to acid side chains of the protein and/or the phosphate groups of the lipids. Iron type 2 is in the 2+ high spin state and is linked to carboxy groups of the protein in a rather unspecific way. Dynamics as measured by Mossbauer spectroscopy show that the purple membrane becomes flexible only above 220 K. At the interface between membrane and bulk water the mobility is comparable to that of proteins with hydrophilic surfaces. The photocycle of Fe 3+-bR is slowed down compared to native bR. 3–5 Fe3+/bR are sufficient to inhibit the photocycle turnover by one order of magnitude. This specific effect is also found with Cr3+, though it is less pronounced. Mössbauer spectra of Fe3+-bR at 4 K reveal that iron nuclei are spin-coupled, indicating their close spatial proximity. It is proposed that iron trinuclear clusters interact with the proton uptake site of bR.
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