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

Abstract

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.

Authors:
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Publication Date:
Research Org.:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
Earth Sciences Division; Life Sciences Division; Physical Biosciences Division
OSTI Identifier:
986219
Report Number(s):
LBNL-3803E-Poster
TRN: US201018%%335
DOE Contract Number:  
DE-AC02-05CH11231
Resource Type:
Technical Report
Resource Relation:
Conference: 110th General Meeting of the American Society for Microbiology, San Diego, CA, May 23-27, 2010
Country of Publication:
United States
Language:
English
Subject:
54; AMMONIUM SULFATES; ANIONS; CHROMATOGRAPHY; DESULFOVIBRIO; FILTRATION; PEPTIDES; PRECIPITATION; PROTEINS; PURIFICATION; TRYPSIN; WATER; tagless purification of water soluble complexes, chromatography

Citation Formats

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 C, Hazen, Terry, Witkowska, H E, Jin, Jiming, Glaeser, Robert, and Biggin, Mark. High Throughput Identification, Purification and Structural Characterization of Water Soluble Protein Complexes in Desulfovibrio vulgaris. United States: N. p., 2010. Web. doi:10.2172/986219.
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 C, Hazen, Terry, Witkowska, H E, Jin, Jiming, Glaeser, Robert, & Biggin, Mark. High Throughput Identification, Purification and Structural Characterization of Water Soluble Protein Complexes in Desulfovibrio vulgaris. United States. https://doi.org/10.2172/986219
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 C, Hazen, Terry, Witkowska, H E, Jin, Jiming, Glaeser, Robert, and Biggin, Mark. 2010. "High Throughput Identification, Purification and Structural Characterization of Water Soluble Protein Complexes in Desulfovibrio vulgaris". United States. https://doi.org/10.2172/986219. https://www.osti.gov/servlets/purl/986219.
@article{osti_986219,
title = {High Throughput Identification, Purification and Structural Characterization of Water Soluble Protein Complexes in Desulfovibrio vulgaris},
author = {Dong, Ming and Han, Bong-Gyoon and Liu, Hui-Hai and Malik, J and Geller, Jil and Yang, Li and Choi, M and Chandonia, John-Marc and Arbelaez, Pablo and Sterling, H J and Typke, Dieter and Shatsky, Max and Brenner, Steve and Fisher, Susan and Williams, Evan and Szakal, Evelin and Allen, S and Hall, S C and Hazen, Terry and Witkowska, H E and Jin, Jiming and Glaeser, Robert and Biggin, Mark},
abstractNote = {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.},
doi = {10.2172/986219},
url = {https://www.osti.gov/biblio/986219}, journal = {},
number = ,
volume = ,
place = {United States},
year = {Mon May 17 00:00:00 EDT 2010},
month = {Mon May 17 00:00:00 EDT 2010}
}