4-Bromophenacyl bromide specifically inhibits rhoptry secretion during Toxoplasma invasion.
Toxoplasma gondii is a eukaryotic parasite of the phylum Apicomplexa that is able to infect a wide variety of host cells. During its active invasion process it secretes proteins from discrete secretory organelles: the micronemes, rhoptries and dense granules. Although a number of rhoptry proteins have been shown to be involved in important interactions with the host cell, very little is known about the mechanism of secretion of any Toxoplasma protein into the host cell. We used a chemical inhibitor of phospholipase A2s, 4-bromophenacyl bromide (4-BPB), to look at the role of such lipases in the secretion of Toxoplasma proteins. We found that 4-BPB was a potent inhibitor of rhoptry secretion in Toxoplasma invasion. This drug specifically blocked rhoptry secretion but not microneme secretion, thus effectively showing that the two processes can be de-coupled. It affected parasite motility and invasion, but not attachment or egress. Using propargyl- or azido-derivatives of the drug (so-called click chemistry derivatives) and a series of 4-BPB-resistant mutants, we found that the drug has a very large number of target proteins in the parasite that are involved in at least two key steps: invasion and intracellular growth. This potent compound, the modified "click-chemistry" forms of it, and the resistant mutants should serve as useful tools to further study the processes of Toxoplasma early invasion, in general, and rhoptry secretion, in particular.
Ravindran
Sandeep
S
Department of Microbiology and Immunology, Stanford University, Stanford, California, United States of America.
Lodoen
Melissa B
MB
Verhelst
Steven H L
SH
Bogyo
Matthew
M
Boothroyd
John C
JC
D000098:Acetophenones / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D000818:Animals; D002448:Cell Adhesion / Q000187:drug effects; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism; D015638:Cytochalasin D / Q000494:pharmacology; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism*; D009068:Movement / Q000187:drug effects; D009154:Mutation / Q000235:genetics; D010271:Parasites / Q000166:cytology / Q000187:drug effects / Q000254:growth & development / Q000472:pathogenicity; D015800:Protozoan Proteins / Q000378:metabolism; D014122:Toxoplasma / Q000166:cytology / Q000187:drug effects* / Q000254:growth & development / Q000472:pathogenicity*; D014617:Vacuoles / Q000187:drug effects / Q000378:metabolism
D000098:Acetophenones; D008565:Membrane Proteins; D015800:Protozoan Proteins; C506860:RON4 protein, Toxoplasma gondii; C117001:ROP 1 protein, Toxoplasma gondii; D015638:Cytochalasin D; C006821:4-bromophenacyl bromide