Hematin crystallization is the primary mechanism of
heme detoxification in
malaria parasites and the target of the
quinoline class of
antimalarials. Despite numerous studies of
malaria pathophysiology, fundamental questions regarding
hematin growth and inhibition remain. Among them are the identity of the crystallization medium in vivo, aqueous or organic; the mechanism of crystallization, classical or nonclassical; and whether
quinoline antimalarials inhibit crystallization by sequestering
hematin in the
solution, or by blocking surface sites crucial for growth. Here we use time-resolved in situ atomic force microscopy (AFM) and show that the
lipid subphase in the parasite may be a preferred growth medium. We provide, to our knowledge, the first evidence of the molecular mechanisms of
hematin crystallization and inhibition by
chloroquine, a common
quinoline antimalarial drug. AFM observations demonstrate that crystallization strictly follows a classical mechanism wherein new crystal layers are generated by 2D nucleation and grow by the attachment of solute molecules. We identify four classes of surface sites available for binding of potential drugs and propose respective mechanisms of
drug action. Further studies reveal that
chloroquine inhibits
hematin crystallization by binding to molecularly flat {100} surfaces. A 2-μM concentration of
chloroquine fully arrests layer generation and step advancement, which is ∼10(4)× less than
hematin's physiological concentration. Our results suggest that adsorption at specific growth sites may be a general mode of
hemozoin growth inhibition for the
quinoline antimalarials. Because the atomic structures of the identified sites are known, this insight could advance the future design and/or optimization of new
antimalarials.