title: Effective elimination of nucleic acids from bacterial protein samples for optimized blue native polyacrylamide gel electrophoresis.
abstract: Nucleic acids remaining within bacterial protein samples from Streptomyces lividans and Escherichia coli were found to interfere significantly with blue native polyacrylamide gel electrophoresis (BN-PAGE), a technique used frequently for analyzing bacterial protein complexes in proteomics studies. We have used ultracentrifugation and/or precipitation of cell lysates with streptomycin sulfate to eliminate nucleic acids from total and/or membrane protein samples. Nucleic acid-binding proteins were first enriched by precipitation with streptomycin sulfate, and contaminating nucleic acids were then eliminated by precipitation by adding polyethyleneimine. The performance of BN-PAGE was found to be dramatically improved by these sample preparation steps.
authors: Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology, Shanghai Jiaotong University, Shanghai, P. R. China.
authors: Jingdan
authors: J
authors: Liang
authors: Qiule
authors: Q
authors: Niu
authors: Xinping
authors: X
authors: Xu
authors: Yuanming
authors: Y
authors: Luo
authors: Xiufen
authors: X
authors: Zhou
authors: Zixin
authors: Z
authors: Deng
authors: Zhijun
authors: Z
authors: Wang
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry / Q000302:isolation & purification*; D011232:Chemical Precipitation; D004269:DNA, Bacterial / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D004926:Escherichia coli / Q000737:chemistry; D029968:Escherichia coli Proteins / Q000737:chemistry; D008565:Membrane Proteins / Q000737:chemistry; D009696:Nucleic Acids / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D012329:RNA, Bacterial / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D048170:Streptomyces lividans / Q000737:chemistry; D013307:Streptomycin / Q000737:chemistry; D014461:Ultracentrifugation
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D008565:Membrane Proteins; D009696:Nucleic Acids; D012329:RNA, Bacterial; D013307:Streptomycin