کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
2020559 1069188 2013 9 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
A general path for large-scale solubilization of cellular proteins: From membrane receptors to multiprotein complexes
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شیمی
پیش نمایش صفحه اول مقاله
A general path for large-scale solubilization of cellular proteins: From membrane receptors to multiprotein complexes
چکیده انگلیسی

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.


► Efficient, high-throughput method to solubilize aggregated and insoluble proteins.
► Successfully tested in bacterial and eukaryotic expression systems.
► Successfully used for membrane, protoplasmic and multi-protein complexes.
► Protein yields in milligram quantities allow biochemical and industrial applications.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Protein Expression and Purification - Volume 87, Issue 2, February 2013, Pages 111–119
نویسندگان
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