A dual-affinity gene fusion system to express small recombinant proteins in a soluble form: expression and characterization of protein A deletion mutants |
| |
Authors: | Jansson Birger; Palmcrantz Carolina; Uhlen Mathias; Nilsson Bjorn |
| |
Affiliation: | Royal Institute of Technology, Department of Biochemistry and Biotechnology S-100 44 Stockholm, Sweden |
| |
Abstract: | A novel gene fusion system to express and purify small recombinantproteins in Escherichia coli has been constructed. The conceptallows for affinity purification of soluble gene products bysequential albumin- and Zn2+-affinity chromatography. The dual-affinitysystem is well suited for expression of unstable proteins asonly full-length protein is obtained after purification andproteins gain proteolytic stability in the fusion protein. Herewe show that the dual-affinity approach can be used for theexpression of various unstable derivatives of a single IgG-bindingdomain based on staphylococcal protein A. Analysis of the proteolyticstabilities and the IgG-binding properties of the differentmutant proteins suggest that the model for the structure ofan IgG-binding domain must be re-evaluated. |
| |
Keywords: | Escherichia coli/ expression system/ staphylococcal protein A/ deletion mutants |
本文献已被 Oxford 等数据库收录! |