Human ribosomal protein S13: cloning, expression, refolding, and structural stability

✍ Scribed by Alexey Malygin; Natalia Parakhnevitch; Galina Karpova

• Publisher: Elsevier Science
• Year: 2005
• Tongue: English
• Weight: 249 KB
• Volume: 1747
• Category: Article
• ISSN: 1570-9639
• DOI: 10.1016/j.bbapap.2004.10.001

✦ Synopsis

The cDNA of human ribosomal protein S13 was cloned into the expression vector pET-15b. Large-scale production of the recombinant protein was carried out in Escherichia coli cells. Protein accumulated in the form of inclusion bodies was isolated, purified, and refolded by dialysis. The recombinant protein was immunologically reactive, interacting with antiserum against native rpS13. The secondary structure content of the refolded protein was analyzed by means of CD spectroscopy. It was found that 43F5% of amino acids sequence of the protein form a-helices and 11F3% are placed in h-strands that coincides with theoretical predictions. The h-strands seem to be located in the extension regions of the rpS13 and do not have homologuous regions in the structure of rpS15 from Thermus thermophilus, which is a prokaryotic homolog of rpS13. The protein structure is stable at a pH range from 4.0 to 8.0 and at low concentrations of urea (up to 3 M).