Spectroscopic characterization of the heme-binding sites in Plasmodium falciparum histidine-rich protein 2.
Proteolysis of hemoglobin provides an essential nutrient source for the malaria parasite Plasmodium falciparum during the intraerythrocytic stage of the parasite's lifecycle. Detoxification of the liberated heme occurs through a unique heme polymerization pathway, leading to the formation of hemozoin. Heme polymerization has been demonstrated in the presence of P. falciparum histidine-rich protein 2 (PfHRP2) [Sullivan, D. J., Gluzman, I. Y., and Goldberg, D. E. (1996) Science 271, 219-221]; however, the molecular role that PfHRP2 plays in this polymerization is currently unknown. PfHRP2 is a 30 kDa protein composed of several His-His-Ala-His-His-Ala-Ala-Asp repeats and is present in the parasite food vacuole, the site of hemoglobin degradation and heme polymerization. We found that, at pH 7.0, PfHRP2 forms a saturable complex with heme, with a PfHRP2 to heme stoichiometry of 1:50. Spectroscopic characterization of heme binding by electronic absorption, resonance Raman, and EPR has shown that bound hemes share remarkably similar heme environments as >95% of all bound hemes are six-coordinate, low-spin, and bis-histidyl ligated. The PfHRP2-ferric heme complex at pH 5.5 (pH of the food vacuole) has the same heme spin state and coordination as observed at pH 7.0; however, polymerization occurs as heme saturation is approached. Therefore, formation of a PfHRP2-heme complex appears to be a requisite step in the formation of hemozoin.
Choi
C Y
CY
Department of Biological Chemistry, Division of Medicinal Chemistry, Howard Hughes Medical Institute, University of Michigan, Ann Arbor 48109-0606, USA.
Cerda
J F
JF
Chu
H A
HA
Babcock
G T
GT
Marletta
M A
MA
D000818:Animals; D001665:Binding Sites; D002248:Carbon Monoxide / Q000737:chemistry; D004578:Electron Spin Resonance Spectroscopy; D005290:Ferric Compounds / Q000737:chemistry; D005296:Ferrous Compounds / Q000737:chemistry; D006418:Heme / Q000737:chemistry / Q000378:metabolism*; D006639:Histidine / Q000378:metabolism*; D010963:Plasmodium falciparum; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D015800:Protozoan Proteins / Q000737:chemistry / Q000378:metabolism*; D013053:Spectrophotometry / Q000379:methods; D013059:Spectrum Analysis, Raman; D000099053:Histidine-Rich Glycoprotein
D005290:Ferric Compounds; D005296:Ferrous Compounds; D011506:Proteins; D015800:Protozoan Proteins; D000099053:Histidine-Rich Glycoprotein; D006418:Heme; D006639:Histidine; D002248:Carbon Monoxide