Growth of Large Hematin Crystals in Biomimetic Solutions

Hematin crystallization is an essential component of the physiology of malaria parasites. Several antimalarial drugs are believed to inhibit crystallization and expose the parasites to toxic soluble hematin. Hence, understanding the mechanisms of hematin crystal growth and inhibition is crucial for the design of new drugs. A major obstacle to microscopic, spectroscopic, and crystallographic studies of hematin crystallization has been the unavailability of large hematin crystals grown under conditions representative of the parasite anatomy. We have developed a biomimetic method to reproducibly grow large hematin crystals reaching 50 pm in length. We imitate the digestive vacuole of Plasmodium falciparum and employ a two-phase solution of octanol and citric buffer. The nucleation of seeds is enhanced at the interface between the aqueous and organic phases, where an ordered layer of octanol molecules is known to serve as substrate for nucleation. The seeds are transferred to hematin-saturated octanol in contact with citric buffer. We show that the crystals grow in the octanol layer, while the buffer supplies hydrogen ions needed for bonds that link the hematin molecules in the crystal. The availability of large hematin crystals opens new avenues for studies of hematin detoxification of malaria parasites in host erythrocytes.