Chemical Biology Tools To Investigate Malaria Parasites

Abstract

Parasitic diseases like malaria tropica have been shaping human evolution and history since the beginning of mankind. After infection, the response of the human host ranges from asymptomatic to severe and may culminate in death. Therefore, proper examination of the parasite's biology is pivotal to deciphering unique molecular, biochemical and cell biological processes, which in turn ensure the identification of treatment strategies, such as potent drug targets and vaccine candidates. However, implementing molecular biology methods for genetic manipulation proves to be difficult for many parasite model organisms. The development of fast and straightforward applicable alternatives, for instance small‐molecule probes from the field of chemical biology, is essential. In this review, we will recapitulate the highlights of previous molecular and chemical biology approaches that have already created insight and understanding of the malaria parasite Plasmodium falciparum. We discuss current developments from the field of chemical biology and explore how their application could advance research into this parasite in the future. We anticipate that the described approaches will help to close knowledge gaps in the biology of P. falciparum and we hope that researchers will be inspired to use these methods to gain knowledge – with the aim of ending this devastating disease.

Novel secretory organelles of parasite origin ‐ at the center of host‐parasite interaction

“…[199][200][201] Another challenge is the genetic manipulation of the different human-pathogenic Plasmodium species and other parasites replicating within RBC. [199,202,203] Transient transfection is very commonly used in research based on certain mammalian cell lines and this technique has also been successfully applied to investigate P. falciparum. However, the well-established, but time-consuming, creation of stable parasite cell lines is usually the method of choice.…”

“…Ready-to-use small molecular probes (e.g., for click chemistry) from the field of chemical biology permit a fast assessment of proteins at every developmental stage of P. falciparum using different experimental approaches. [202,213,214] The combination of chemical probes and correlative light and electron microscopy (CLEM) could potentially help in the assessment of protein localization and organelle morphology. [215,216] Moreover, molecular probes allow the investigation of other parasites that reside within RBC or reticulocytes (Table 1) and are difficult to assess at the moment.…”

“…Ready‐to‐use small molecular probes (e.g., for click chemistry) from the field of chemical biology permit a fast assessment of proteins at every developmental stage of P. falciparum using different experimental approaches. [ 202,213,214 ]…”

Chemo‐proteomics in antimalarial target identification and engagement

“…The copper‐mediated CuAAC reaction uses terminal azide and alkyne functionalities to form a 1,2,3‐triazole linkage based on the Huisgen Cycloaddition (Figure 2A). However, the use of cytotoxic copper reagents can be undesirable and are therefore not applicable for use in live Plasmodium 113 . Therefore, copper‐free methods have come into prominence for this purpose.…”

Plasmodium schizogony, a chronology of the parasite’s cell cycle in the blood stage

“…The list of live–cell markers for schizogony is, however, steadily growing ( Table 1 ). As the generation of transgenic parasite lines is still a rate-limiting step, the use of state-of-the-art live–cell dyes is an important tool [ 118 ]. Due to their excellent background-to-noise ratio, fluorogenic dyes have significantly contributed to this progress [ 119 , 120 ], while classical dyes like the membrane label BODIPY still have their utility [ 46 , 116 ].…”