The Malaria Pigment Hemozoin Comprises at Most Four Different Isomer Units in Two Crystalline Models: Chiral as Based on a Biochemical Hypothesis or Centrosymmetric Made of Enantiomorphous Sectors

TY - JOUR

T1 - The Malaria Pigment Hemozoin Comprises at Most Four Different Isomer Units in Two Crystalline Models

T2 - Chiral as Based on a Biochemical Hypothesis or Centrosymmetric Made of Enantiomorphous Sectors

AU - Straasø, Tine

AU - Marom, Noa

AU - Solomonov, Inna

AU - Barfod, Lea Klingenberg

AU - Burghammer, Manfred

AU - Feidenhans'l, Robert Krarup

AU - Als-Nielsen, Jens

AU - Leiserowitz, Leslie

PY - 2014/4/2

Y1 - 2014/4/2

N2 - Hemozoin is a crystalline byproduct formed upon hemoglobin digestion in Plasmodium-infected blood cells. Based on X-ray powder diffraction (XRPD), hemozoin and its synthetic analogue β-hematin are very similar in structure, consisting of cyclic dimers (cd) of ferriprotoporphyrin IX [Fe(3+)-PPIX] molecules coordinated via Fe-O(propionate) bonds. Enantiofacial symmetry of Fe(3+)-PPIX implies formation of four different stereoisomeric dimers, two centrosymmetric (1̄), labeled cd1̄1 and cd1̄2, and two enantiomeric, cd2(+) and cd2(-), in which the Fe(3+)PPIX moieties are related by pseudo-2-fold symmetry. Only the cd1̄1 stereoisomer was reported as the repeat unit in the initial structural elucidation of β-hematin and refinement of hemozoin. Our recent study of β-hematin, employing a combination of XRPD and density functional theory (DFT), revealed besides the published phase, characterized in terms of a disordered cd1̄1/cd2(±) mixture, which is diffractionally equivalent to a cd1̄1/cd1̄2 mixture, a minor phase considered to comprise mainly cd1̄2 dimers. As a consequence single-phase β-hematin powders were recently reanalyzed in terms of a cd1̄1/cd1̄2 mixture, yielding an average occupancy ≅75:25. Here, we present evidence enhancing the biphase model of β-hematin. The primary focus is on a reexamination of the hemozoin structure in light of a biochemically based dimerization mechanism that we recently hypothesized. We suggest that upon hemoglobin degradation, the heme byproduct retains the O2 molecule bound to Fe on the Re side of the heme until Fe-O(propionate) coordination between such heme molecules occurs across their unbound Si sides yielding the cd2(+) dimer. We report Rietveld refinement of the hemozoin structure using data measured on an all-in-vacuum powder diffractometer assuming the following models: cd1̄1, cd1̄2, cd2(+), and the two mixtures cd1̄1/ cd1̄2 and cd1̄1/cd2(+). The best figures of merit were obtained for the mixture cd1̄1/cd2(+) with a 50:50 occupancy, followed by the cd1̄1/cd1̄2 mixture with an occupancy ≅75:25, which we interpret as a structure that comprises the cd1̄1, cd2(+), cd2(-), and cd1̄2 isomers with occupancies ≅58:17:17:8. In this model system, the cd1̄1 "host" molecule is uniformly distributed throughout the crystal, whereas the enantiomeric molecules cd2(+) and cd2(-) are preferentially occluded in different crystalline sectors, which are thus enantiomorphous, related by overall centrosymmetric symmetry. Various arguments appear to favor the 50:50 cd1̄1/cd2(+) mixture, namely, a hemozoin crystal of overall chiral symmetry, consistent with our hypothesis. However, we cannot overrule the alternative model.

AB - Hemozoin is a crystalline byproduct formed upon hemoglobin digestion in Plasmodium-infected blood cells. Based on X-ray powder diffraction (XRPD), hemozoin and its synthetic analogue β-hematin are very similar in structure, consisting of cyclic dimers (cd) of ferriprotoporphyrin IX [Fe(3+)-PPIX] molecules coordinated via Fe-O(propionate) bonds. Enantiofacial symmetry of Fe(3+)-PPIX implies formation of four different stereoisomeric dimers, two centrosymmetric (1̄), labeled cd1̄1 and cd1̄2, and two enantiomeric, cd2(+) and cd2(-), in which the Fe(3+)PPIX moieties are related by pseudo-2-fold symmetry. Only the cd1̄1 stereoisomer was reported as the repeat unit in the initial structural elucidation of β-hematin and refinement of hemozoin. Our recent study of β-hematin, employing a combination of XRPD and density functional theory (DFT), revealed besides the published phase, characterized in terms of a disordered cd1̄1/cd2(±) mixture, which is diffractionally equivalent to a cd1̄1/cd1̄2 mixture, a minor phase considered to comprise mainly cd1̄2 dimers. As a consequence single-phase β-hematin powders were recently reanalyzed in terms of a cd1̄1/cd1̄2 mixture, yielding an average occupancy ≅75:25. Here, we present evidence enhancing the biphase model of β-hematin. The primary focus is on a reexamination of the hemozoin structure in light of a biochemically based dimerization mechanism that we recently hypothesized. We suggest that upon hemoglobin degradation, the heme byproduct retains the O2 molecule bound to Fe on the Re side of the heme until Fe-O(propionate) coordination between such heme molecules occurs across their unbound Si sides yielding the cd2(+) dimer. We report Rietveld refinement of the hemozoin structure using data measured on an all-in-vacuum powder diffractometer assuming the following models: cd1̄1, cd1̄2, cd2(+), and the two mixtures cd1̄1/ cd1̄2 and cd1̄1/cd2(+). The best figures of merit were obtained for the mixture cd1̄1/cd2(+) with a 50:50 occupancy, followed by the cd1̄1/cd1̄2 mixture with an occupancy ≅75:25, which we interpret as a structure that comprises the cd1̄1, cd2(+), cd2(-), and cd1̄2 isomers with occupancies ≅58:17:17:8. In this model system, the cd1̄1 "host" molecule is uniformly distributed throughout the crystal, whereas the enantiomeric molecules cd2(+) and cd2(-) are preferentially occluded in different crystalline sectors, which are thus enantiomorphous, related by overall centrosymmetric symmetry. Various arguments appear to favor the 50:50 cd1̄1/cd2(+) mixture, namely, a hemozoin crystal of overall chiral symmetry, consistent with our hypothesis. However, we cannot overrule the alternative model.

U2 - 10.1021/cg401151f

DO - 10.1021/cg401151f

M3 - Journal article

SN - 1528-7483

VL - 14

SP - 1543

EP - 1554

JO - Crystal Growth & Design

JF - Crystal Growth & Design

IS - 4

ER -