High Throughput Identification, Purification and Structural Characterization of Water Soluble Protein Complexes in Desulfovibrio vulgaris

Dong,, Ming ; Han, Bong-Gyoon ; Liu, Hui-Hai ; Malik, J. ; Geller, Jil ; Yang, Li ; Choi, M. ; Chandonia, John-Marc ; Arbelaez, Pablo ; Sterling, H. J. ; Typke, Dieter ; Shatsky, Max ; Brenner, Steve ; Fisher, Susan ; Williams, Evan ; Szakal, Evelin ; Allen, S. ; Hall, S.

关键词: 54;    AMMONIUM SULFATES;    ANIONS;    CHROMATOGRAPHY;    DESULFOVIBRIO;    FILTRATION;    PEPTIDES;    PRECIPITATION;    PROTEINS;    PURIFICATION;    TRYPSIN;    WATER tagless purification of water soluble complexes;    chromatography;   

DOI  :  10.2172/986219
RP-ID  :  LBNL-3803E-Poster
PID  :  OSTI ID: 986219
Others  :  TRN: US201018%%335

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来源: SciTech Connect

【 摘 要 】

Our scheme for the tagless purification of water soluble complexes. 10 g of protein from a crude bacterial extract is first fractionated by ammonium sulfate precipitation and then by a series of chromatographic steps: anion exchange (IEX), hydrophobic interaction (HIC), and finally size exclusion (Gel Filtration). Fractions from the last chromatography step are trypsin digested and peptides labeled with iTRAQ reagents to allow multiplexing and quantitation during mass spectrometric analysis. Elution profiles of identified proteins are then subjected to clustering analysis.