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Full-Text Articles in Physical Sciences and Mathematics

Detection Of Dityrosine In Apoferritin, Stephen E. Bialkowski Jan 1995

Detection Of Dityrosine In Apoferritin, Stephen E. Bialkowski

Stephen E. Bialkowski

Laser-excited fluorescence spectroscopy was adopted for dityrosine detection in horse spleen apoferritin. Dityrosine was identified in horse spleen apoferritin. Fractionation of acid-hydrolyzed horse spleen apoferritin showed fluorescence attributed to dityrosine, and other compounds, excited by 325-nm laser radiation. The other fluorescent compounds resulted in a broad fluorescence emission spectrum of horse spleen apoferritin. The broad emission band overlapped with the fluorescence emission spectrum of pure dityrosine. Fractionation of horse spleen apoferritin acid hydrolysate prior to laser-excited fluorescence detection was necessary for dityrosine quantitative analysis. The concentration of dityrosine in horse spleen apoferritin was 2.5% (mol/mol), estimated from the method of …


Laser Excited Fluorescence Of Dityrosine, Sahar F. Mahmoud, Stephen E. Bialkowski Jan 1995

Laser Excited Fluorescence Of Dityrosine, Sahar F. Mahmoud, Stephen E. Bialkowski

Stephen E. Bialkowski

In this research, laser-excited fluorescence was examined for sensitive detection of aqueous dityrosine. Samples were excited with a 6.3-mW, 325-nm helium-cadmium laser focused into a small volume-fluorescence cell with a 10-cm lens. The resulting fluorescence emission was collected perpendicular to the excitation and detected with two different schemes. An optical bandpass filter was used with a photomultiplier tube for sensitive quantitative measurement, while a photodiode array detector was used in conjunction with a spectrograph for qualitative characterization of fluorescence emission spectra. Dityrosine detection on the order of 2 × 10-11 M was obtained with the use of the photomultiplier …


Formation Of O-Tyrosine And Dityrosine In Proteins During Radiolytic And Metal-Catalyzed Oxidation, Thomas G. Huggins, Mary C. Wells-Knecht, Nicholas A. Detorie, John W. Baynes, Suzanna R. Thorpe Jun 1993

Formation Of O-Tyrosine And Dityrosine In Proteins During Radiolytic And Metal-Catalyzed Oxidation, Thomas G. Huggins, Mary C. Wells-Knecht, Nicholas A. Detorie, John W. Baynes, Suzanna R. Thorpe

Faculty Publications

To evaluate their usefulness as chemical indicators of cumulative oxidative damage to proteins, we studied the kinetics and extent of formation of ortho-tyrosine (0-Tyr), dityrosine (DT), and dityrosine-like fluorescence (Ex = 3 17 nm, E,,, = 407 nm) in the model proteins RNase and lysozyme exposed to radiolytic and metalcatalyzed (H20z/Cu2+) oxidation (MCO). Although there were protein-dependent differences, o-Tyr, DT, and fluorescence increased coordinately during oxidation of the proteins in both oxidation systems. The contribution of DT to total dityrosine-like fluorescence in oxidized proteins varied from 2-10070, depending on the protein, type of oxidation, and extent of oxidative damage. In …


Oxidized Amino Acids In Lens Protein With Age: Measurement Of O-Tyrosine And Dityrosine In The Aging Human Lens, Mary C. Wells-Knect, Thomas G. Huggins, Daniel G. Dyer, Suzanne R. Thorpe, John W. Baynes Jun 1993

Oxidized Amino Acids In Lens Protein With Age: Measurement Of O-Tyrosine And Dityrosine In The Aging Human Lens, Mary C. Wells-Knect, Thomas G. Huggins, Daniel G. Dyer, Suzanne R. Thorpe, John W. Baynes

Faculty Publications

The concentrations of ortho-tyrosine (o-Tyr) and dityrosine (DT) were measured in noncataractous human lenses in order to assess the role of proteinoxidation reactions in the aging of lens proteins. The measurements were conducted by selected ion monitoring-gas chromatography/mass spectrometry using deuterium-labeled internal standards, which provided both high sensitivity and specificity for the quantitation of o-Tyr and DT. Between ages 1 and 78 years, the o-Tyr concentration in lens proteins varied from 0.3 to 0.9 mmol of o-Tyr/mol of Phe (n = 19), while DT ranged from 1 to 3 mumol of DT/mol of Tyr (n = 30). There were no …