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Expression of porin from Rhodopseudomonas blastica in Escherichia coli inclusion bodies and folding into exact native structure
Author(s) -
Schmid Benedikt,
Krömer Markus,
Schulz Georg E.
Publication year - 1996
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(96)00080-4
Subject(s) - porin , escherichia coli , folding (dsp implementation) , bacterial outer membrane , inclusion bodies , chemistry , microbiology and biotechnology , inclusion (mineral) , protein folding , biology , biochemistry , gene , mineralogy , electrical engineering , engineering
The homotrimeric membrane channel porin from Rhodopseudomonas blastica was expressed without signal sequence in Escherichia coli . The protein assembled in inclusion bodies in the cytosol, from which it could be recovered using urea and detergents. After purification by anion‐exchange chromatography, the protein crystallized under wild‐type conditions. The X‐ray structure was determined at 2.2 Å resolution, and a comparison with the known wild‐type structure showed that the recombinant porin is identical at the atomic level. The method yields porin and designed mutants thereof in 100 mg amounts, allowing for detailed functional and mechanistic studies.