Capillary sieving electrophoresis-micellar electrokinetic chromatography fully automated two-dimensional capillary electrophoresis analysis of Deinococcus radiodurans protein homogenate

被引:75
|
作者
Michels, DA
Hu, S
Dambrowitz, KA
Eggertson, MJ
Lauterbach, K
Dovichi, NJ [1 ]
机构
[1] Univ Washington, Dept Chem, Seattle, WA 98915 USA
[2] Univ Calif Los Angeles, Sch Dent, Los Angeles, CA 90024 USA
关键词
capillary sieving electrophoresis; Deinococcus radiodurans; micellar electrokinetic capillary chromatography;
D O I
10.1002/elps.200405939
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
We report the one- and two-dimensional (1-D and 2-D) capillary electrophoresis separation of Deinococcus radiodurans protein homogenate. Proteins are labeled with the fluorogenic reagent 3-(2-furoyl)quinoline-2-carboxaldehyde (FQ), which reacts with 2 lysine residues and creates a highly fluorescent product. Detection is by laser-induced fluorescence. 1-D capillary sieving electrophoresis (CSE) produces over 150 000 plates and micellar electrokinetic capillary chromatography (MEKC) produces over 900000 plates for components in a D. radiodurans protein homogenate. In a 2-D separation, proteins are first separated by CSE. Fractions are repetitively transferred to a second capillary for further separation based on MEKC. The 2-D separation has a similar to550 spot capacity. Over 150 components are partially resolved from the homogenate. Resolution is limited in the first dimension by diffusion of proteins during the long separation period and in the second dimension by the combination of a long fraction-transfer time and short separation period.
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页码:3098 / 3105
页数:8
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