Theory, design, and characterization of a microdialysis flow cell for Raman spectroscopy

Biophysical Journal
R Tuma, G J Thomas

Abstract

The theory, design, and application of a dialysis flow cell for Raman spectroscopy are described. The flow cell permits rapid collection of Raman spectra concurrent with the efflux of small solute molecules or ions into a solution of macromolecules and is well suited to acquisition of data during hydrogen-isotope exchange reactions of biological molecules. Kinetic parameters of the device are described by a diffusion model, which accounts satisfactorily for the observed rates of efflux of deuterium oxide (K2H = 0.30 min-1), calcium ions (KCa = 0.10 min-1) and EGTA (KEGTA = 0.07 min-1). Application to the kinetics of glutamate protonation in a peptide copolymer [poly(Glu, Lys, Tyr)] shows that pH-titration rates as high as 3.3 pH units/min can be monitored. It is also shown that one can extract first-order hydrogen-isotope exchange rate constants from measured second-order exchanges by taking into account the rate of entry of 2H2O effluent into the bulk H2O solution. Deuterium exchanges of the single-stranded polyribonucleotides poly(rA) and poly(rU) and of the double-stranded RNA genome from bacteriophage phi 6 have been investigated. The measured nucleotide base exchange rates are comparable with those determined previously by...Continue Reading

Citations

Mar 31, 2006·Analytical and Bioanalytical Chemistry·Arantxa Rodríguez-CasadoPedro Carmona
Dec 4, 2008·Analytical and Bioanalytical Chemistry·Pedro CarmonaMarina Molina
Jul 20, 1999·Annual Review of Biophysics and Biomolecular Structure·G J Thomas
Aug 26, 1998·Proceedings of the National Academy of Sciences of the United States of America·R TumaG J Thomas
Nov 14, 2015·The Journal of Physical Chemistry. B·Gregory V BarnettChristopher J Roberts
Jul 29, 1998·Journal of Molecular Biology·R TumaP E Prevelige
Aug 12, 1998·Journal of Molecular Biology·L Laporte, G J Thomas
Jun 27, 1998·Journal of Molecular Biology·J T JuutiG J Thomas
May 5, 2004·Biochemistry·James M BenevidesGeorge J Thomas

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