Expression and purification of anthrax toxin protective antigen from Escherichia coli

Protein Expression and Purification
M SharmaY Singh

Abstract

Anthrax toxin consists of three separate proteins, protective antigen (PA), lethal factor (LF), and edema factor (EF). PA binds to the receptor on mammalian cells and facilitates translocation of EF or LF into the cytosol. PA is the primary component of several anthrax vaccines. In this study we expressed and purified PA from Escherichia coli. The purification of PA from E. coli was possible after transporting the protein into the periplasmic space using the outer membrane protein A signal sequence. The purification involved sequential chromatography through hydroxyapatite, DEAE Sepharose CL-4B, followed by Sephadex G-100. The typical yield of purified PA from this procedure was 500 microg/liter. PA expressed and purified from E. coli was similar to the PA purified from Bacillus anthracis in its ability to lyse a macrophage cell line (J774A.1). The present results suggest that a signal sequence is required for the efficient translocation of PA into E. coli periplasmic space.

Citations

Jul 26, 2008·Applied Biochemistry and Biotechnology·Fatemeh VahediMahmoud Mahmoudi
Mar 12, 2016·Molecular Biology International·Nagendra SuryanarayanaUrmil Tuteja
Jun 27, 2006·Journal of Biotechnology·Hsin-Hou ChangDer-Shan Sun
Jul 13, 2019·Expert Review of Vaccines·Olga A KondakovaOlga V Karpova
Feb 7, 1998·Infection and Immunity·P GuptaR Bhatnagar
Aug 4, 2001·Biochemical and Biophysical Research Communications·N AhujaR Bhatnagar
Jul 30, 1999·Protein Expression and Purification·P GuptaR Bhatnagar
Sep 14, 2001·Biochemical and Biophysical Research Communications·N AhujaR Bhatnagar
Jun 25, 2015·Microbiology Spectrum·Susan WelkosChristopher Cote
Jul 26, 2003·Protein Expression and Purification·Joungmok KimMoon-Young Yoon
Aug 2, 2003·Biochemical and Biophysical Research Communications·Nidhi AhujaRakesh Bhatnagar
Oct 13, 2004·Protein Expression and Purification·Michael W LairdWilliam Gwinn

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