Expression and purification of anthrax toxin protective antigen from Escherichia coli.

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.

journal_name

Protein Expr Purif

authors

Sharma M,Swain PK,Chopra AP,Chaudhary VK,Singh Y

doi

10.1006/prep.1996.0005

subject

Has Abstract

pub_date

1996-02-01 00:00:00

pages

33-8

issue

1

eissn

1046-5928

issn

1096-0279

pii

S1046-5928(96)90005-4

journal_volume

7

pub_type

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