Analysis of major protein-protein and protein-metal complexes of erythrocytes directly from cell lysate utilizing capillary electrophoresis mass spectrometry

Analytical Chemistry
An Nguyen, M Moini

Abstract

The separation and detection of the major protein-protein and protein-metal complexes of erythrocytes directly from cell lysate under native conditions has been accomplished for the first time using capillary electrophoresis electrospray ionization-mass spectrometry (CE/ESI-MS). All three major protein-protein and protein-metal complexes in human red blood cells (RBCs) with a concentration dynamic range of approximately 3 orders of magnitude were successfully detected. Intact complexes detected in lysed RBCs included carbonic anhydrase II (CAII-Zn at approximately 0.8 amol/cell) complexed with its zinc cofactor, carbonic anhydrase I (CAI-Zn at approximately 7 amol/cell) complexed with its zinc cofactor, and hemoglobin A (Hb-tetramer at approximately 450 amol/cell)a tetramer formed by two alpha-beta-subunits and four heme groups. The average molecular weights measured for these complexes were consistent with their theoretical values within 0.01% mass accuracy. The use of Polybrene as a self-coating reagent in conjunction with ammonium acetate at pH approximately 7.4, narrow capillary for high separation efficiency, and forward polarity CE to avoid acid production at the tip of the capillary were overriding experimental factors f...Continue Reading

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Citations

Feb 4, 2012·Analytical and Bioanalytical Chemistry·James P BarnettClaudia A Blindauer
Feb 10, 2012·Journal of the Royal Society, Interface·Gillian R Hilton, Justin L P Benesch
Aug 11, 2011·Analytical Sciences : the International Journal of the Japan Society for Analytical Chemistry·Mio KawaiKazuichi Hayakawa
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Oct 18, 2016·Analytical Chemistry·Gongyu LiGuangming Huang
Apr 11, 2009·Journal of Separation Science·Bryan R Fonslow, John R Yates
Dec 8, 2009·Electrophoresis·Andrei R Timerbaev
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May 26, 2010·Analytical Chemistry·Nicholas W FrostMichael T Bowser
Jun 16, 2009·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Farid E Ahmed

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