• Home
  • Search Results
  • Anthrax toxin requires ZDHHC5-mediated palmitoylation of its surface-processing host enzymes.

Anthrax toxin requires ZDHHC5-mediated palmitoylation of its surface-processing host enzymes.

Proceedings of the National Academy of Sciences of the United States of America (2019-01-06)
Oksana A Sergeeva, F Gisou van der Goot
ABSTRACT

The protein acyl transferase ZDHHC5 was recently proposed to regulate trafficking in the endocytic pathway. Therefore, we explored the function of this enzyme in controlling the action of bacterial toxins. We found that ZDHHC5 activity is required for two very different toxins: the anthrax lethal toxin and the pore-forming toxin aerolysin. Both of these toxins have precursor forms, the protoxins, which can use the proprotein convertases Furin and PC7 for activation. We show that ZDHHC5 indeed affects the processing of the protoxins to their active forms. We found that Furin and PC7 can both be S-palmitoylated and are substrates of ZDHHC5. The impact of ZDHHC5 on Furin/PC7-mediated anthrax toxin cleavage is dual, having an indirect and a direct component. First, ZDHHC5 affects the homeostasis and trafficking of a subset of cellular proteins, including Furin and PC7, presumably by affecting the endocytic/recycling pathway. Second, while not inhibiting the protease activity per se, ZDHHC5-mediated Furin/PC7 palmitoylation is required for the cleavage of the anthrax toxin. Finally, we show that palmitoylation of Furin and PC7 promotes their association with plasma membrane microdomains. Both the receptor-bound toxin and the convertases are of very low abundance at the cell surface. Their encounter is unlikely on reasonable time scales. This work indicates that palmitoylation drives their encounter in specific domains, allowing processing and thereby intoxication of the cell.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Monoclonal ANTI-FLAG® M2 antibody produced in mouse, 1 mg/mL, clone M2, affinity isolated antibody, buffered aqueous solution (50% glycerol, 10 mM sodium phosphate, and 150 mM NaCl, pH 7.4)
Sigma-Aldrich
Minimum Essential Medium Eagle, With Earle′s salts, L-glutamine and sodium bicarbonate, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
N-Hydroxysulfosuccinimide sodium salt, ≥98% (HPLC)
Sigma-Aldrich
Monoclonal Anti-α-Tubulin antibody produced in mouse, ascites fluid, clone B-5-1-2
Sigma-Aldrich
Anti-GAPDH antibody, Mouse monoclonal, clone GAPDH-71.1, purified from hybridoma cell culture
Millipore
Streptavidin−Agarose from Streptomyces avidinii, buffered aqueous suspension
Millipore
Anti-V5 Agarose Affinity Gel antibody produced in mouse, purified immunoglobulin, clone V5-10
Sigma-Aldrich
Benzamidine, ≥95.0%
Sigma-Aldrich
Anti-Goat IgG (whole molecule)–Peroxidase antibody produced in rabbit, affinity isolated antibody, buffered aqueous solution
Sigma-Aldrich
Anti-Chicken IgY (IgG) (whole molecule)−Peroxidase antibody produced in rabbit, affinity isolated antibody, buffered aqueous solution
Sigma-Aldrich
Glasgow Minimum Essential Medium, With L-glutamine, without tryptose phosphate broth and sodium bicarbonate, powder, suitable for cell culture
Sigma-Aldrich
Anti-ZDHHC5 antibody produced in rabbit, Prestige Antibodies® Powered by Atlas Antibodies, affinity isolated antibody, buffered aqueous glycerol solution
Sigma-Aldrich
Anti-TEM8/Anthrax Toxin Receptor 1 antibody produced in goat, affinity isolated antibody, buffered aqueous solution