Abstract：
:Expression of recombinant proteins in bacterial or eukaryotic systems often results in aggregation rendering them unavailable for biochemical or structural studies. Protein aggregation is a costly problem for biomedical research. It forces research laboratories and the biomedical industry to search for alternative, more soluble, non-human proteins and limits the number of potential "druggable" targets. In this study we present a highly reproducible protocol that introduces the systematic use of an extensive number of detergents to solubilize aggregated proteins expressed in bacterial and eukaryotic systems. We validate the usefulness of this protocol by solubilizing traditionally difficult human protein targets to milligram quantities and confirm their biological activity. We use this method to solubilize monomeric or multimeric components of multi-protein complexes and demonstrate its efficacy to reconstitute large cellular machines. This protocol works equally well on cytosolic, nuclear and membrane proteins and can be easily adapted to a high throughput format.
journal_name
Protein Expr Purifjournal_title
Protein expression and purificationauthors
Pullara F,Guerrero-Santoro J,Calero M,Zhang Q,Peng Y,Spåhr H,Kornberg GL,Cusimano A,Stevenson HP,Santamaria-Suarez H,Reynolds SL,Brown IS,Monga SP,Van Houten B,Rapić-Otrin V,Calero G,Levine ASdoi
10.1016/j.pep.2012.10.007subject
Has Abstractpub_date
2013-02-01 00:00:00pages
111-9issue
2eissn
1046-5928issn
1096-0279pii
S1046-5928(12)00294-Xjournal_volume
87pub_type
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journal_title：Protein expression and purification
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journal_title：Protein expression and purification
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journal_title：Protein expression and purification
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journal_title：Protein expression and purification
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journal_title：Protein expression and purification
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journal_title：Protein expression and purification
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journal_title：Protein expression and purification
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journal_title：Protein expression and purification
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