Selection of surfactants for cell lysis in chemical cytometry to study protein-DNA interactions

Electrophoresis
Zhulin PangSergey N Krylov

Abstract

Protein-DNA interactions play a defining role in many cellular processes. Studying such interactions at the single-cell level is important and challenging. Here we make the first step toward achieving this goal with chemical cytometry. Chemical cytometry utilizes capillary separation for detailed chemical analyses of single cells. The cell is injected into a capillary, lysed, and its components are analyzed by CE or capillary chromatography with highly sensitive detection. In order to apply chemical cytometry to studies of protein-DNA interactions, cell lysis must not destroy protein-DNA complexes. Surfactants represent the most practical means of cell lysis inside the capillary. This work aimed at finding surfactants and lysis conditions that do not destroy protein-DNA complexes. We studied three groups of surfactants--ionic, zwitterionic, and nonionic--with respect to their ability to lyse the cell membrane without significantly influencing the stability of protein-DNA complexes. Nonequilibrium CE of equilibrium mixtures with surfactants in the equilibrium mixtures and in the run buffer was used to measure the equilibrium constant, K(d), and rate constant, k(off), of protein-DNA complex dissociation. We found that nonionic su...Continue Reading

References

Oct 6, 1989·Science·R T KennedyJ W Jorgenson
Dec 31, 1965·Science·G T Matioli, H B Niewisch
Mar 1, 1994·Journal of Neuroscience Methods·H K KristensenA G Ewing
Mar 3, 1996·Journal of Chromatography. B, Biomedical Applications·Q Xue, E S Yeung
Nov 21, 1998·Analytical Chemistry·C E SimsN L Allbritton
Mar 4, 2000·Analytical Chemistry·S N KrylovN J Dovichi
Jul 11, 2002·Journal of Chromatography. a·Nicole E Baryla, Charles A Lucy
Oct 31, 1953·Nature·J E EDSTROM
Oct 29, 2003·Current Opinion in Chemical Biology·Norman J Dovichi, Shen Hu
Nov 1, 2003·Analytical Chemistry·Maxine A McClainJ Michael Ramsey
Jan 20, 2004·European Journal of Biochemistry·Christian KlammtFrank Bernhard
Feb 28, 2004·Analytical Chemistry·Victor OkhoninSergey N Krylov
Jun 11, 2004·Biophysical Journal·Tomasz Zal, Nicholas R J Gascoigne
Jul 2, 2004·Analytical Chemistry·Kang HuSergey N Krylov
Aug 26, 2004·Proceedings of the National Academy of Sciences of the United States of America·Hongkai WuRichard N Zare
Sep 9, 2004·The Biochemical Journal·Valerică RaicuJames D Friesen
Oct 27, 2004·Journal of Molecular Biology·James J HavranekDavid Baker
Mar 3, 2005·Journal of the American Chemical Society·Maxim BerezovskiSergey N Krylov

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Citations

Dec 22, 2009·Integrative Biology : Quantitative Biosciences From Nano to Macro·Jungkyu KimBruce K Gale
Apr 23, 2008·Journal of the Royal Society, Interface·Robert B Brown, Julie Audet
Jul 19, 2008·Annual Review of Analytical Chemistry·Laura M BorlandNancy L Allbritton
May 31, 2011·Journal of Biomedical Science·Jasmine P DevadhasanJeongho An
Jun 12, 2009·Electrophoresis·Niels H H Heegaard
Dec 7, 2007·Electrophoresis·Vladislav Dolník
Jan 25, 2007·Electrophoresis·Sergey N Krylov
Apr 4, 2008·Electrophoresis·Hui YanHongkai Wu
Oct 12, 2013·Chemical Biology & Drug Design·Łukasz PałkowskiRoman Słowiński
Dec 7, 2016·Biomedical Microdevices·Aishah PrastowoEdmond J Walsh
Jun 19, 2008·Journal of the American Chemical Society·Maxim V BerezovskiSergey N Krylov
May 17, 2021·Food Research International·Xiaoying HanYing Mu
Jul 29, 2017·ACS Sensors·Salvatore Petralia, Sabrina Conoci
Jun 15, 2007·Analytical Chemistry·Colin D WhitmoreNorman J Dovichi

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