Affinity chromatography and capillary electrophoresis for analysis of the yeast ribosomal proteins

BMB Reports

Keith R. Willison^2¹  David R. Klug^1²  Miriam S. Goyder^1³ 

[1] Institute of Cancer Research, Chester Beatty Laboratories, Section of Cell and Molecular Biology, London, UK SW3 6JB^2;Institute of Chemical and Biochemical Engineering, ETH Z?rich, 8093 Z?rich, Switzerland^3;The Single Cell Proteomics Group, Institute of Chemical Biology, Department of Chemistry, Imperial College London, South Kensington, London, UK SW72AZ^1

关键词: Affinity chromatography;   

DOI  :  

学科分类：生物化学/生物物理

来源: Korean Society for Biochemistry and Molecular Biology

【 摘 要 】

We present a top down separation platform for yeast ribosomal proteins using affinity chromatography and capillary electrophoresis which is designed to allow deposition of proteins onto a substrate. FLAG tagged ribosomes were affinity purified, and rRNA acid precipitation was performed on the ribosomes followed by capillary electrophoresis to separate the ribosomal proteins. Over 26 peaks were detected with excellent reproducibility (<0.5% RSD migration time). This is the first reported separation of eukaryotic ribosomal proteins using capillary electrophoresis. The two stages in this workflow, affinity chromatography and capillary electrophoresis, share the advantages that they are fast, flexible and have small sample requirements in comparison to more commonly used techniques. This method is a remarkably quick route from cell to separation that has the potential to be coupled to high throughput readout platforms for studies of the ribosomal proteome.

【 授权许可】

Unknown