title: Characterization of Precursor PfHsp60 in Plasmodium falciparum Cytosol during Its Asexual Development in Human Erythrocytes.
abstract: Mitochondrial heat shock protein 60 (Hsp60) is a nuclear encoded gene product that gets post-translationally translocated into the mitochondria. Using multiple approaches such as immunofluorescence experiments, isoelectric point analysis with two-dimensional gel electrophoresis, and mass spectrometric identification of the signal peptide, we show that Hsp60 from Plasmodium falciparum (PfHsp60) accumulates in the parasite cytoplasm during the ring, trophozoite, and schizont stages of parasite development before being imported into the parasite mitochondria. Using co-immunoprecipitation experiments with antibodies specific to cytoplasmic PfHsp90, PfHsp70-1, and PfHsp60, we show association of precursor PfHsp60 with cytoplasmic chaperone machinery. Metabolic labeling involving pulse and chase indicates translocation of the precursor pool into the parasite mitochondrion during chase. Analysis of results obtained with Geldanamycin treatment confirmed precursor PfHsp60 to be one of the clients for PfHsp90. Cytosolic chaperones bind precursor PfHsp60 prior to its import into the mitochondrion of the parasite. Our data suggests an inefficient co-ordination in the synthesis and translocation of mitochondrial PfHsp60 during asexual growth of malaria parasite in human erythrocytes.
authors: Department of Physics, Indian Institute of Science, Bangalore, Karnataka, India; Department of Biochemistry, Indian Institute of Science, Bangalore, Karnataka, India.
authors: P
authors: P
authors: Padma Priya
authors: Department of Biochemistry, Indian Institute of Science, Bangalore, Karnataka, India.
authors: Manish
authors: M
authors: Grover
authors: Department of Biochemistry, Indian Institute of Science, Bangalore, Karnataka, India.
authors: Utpal S
authors: US
authors: Tatu
authors: Department of Physics, Indian Institute of Science, Bangalore, Karnataka, India.
authors: Vasant
authors: V
authors: Natarajan
mesh_terms: D016227:Benzoquinones / Q000494:pharmacology; D018834:Chaperonin 60 / Q000235:genetics / Q000378:metabolism*; D004912:Erythrocytes / Q000469:parasitology*; D006801:Humans; D047029:Lactams, Macrocyclic / Q000494:pharmacology; D024101:Mitochondrial Proteins / Q000235:genetics / Q000378:metabolism*; D010963:Plasmodium falciparum / Q000235:genetics / Q000378:metabolism*; D011498:Protein Precursors / Q000235:genetics / Q000378:metabolism*; D021382:Protein Sorting Signals / Q000235:genetics; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D016227:Benzoquinones; D018834:Chaperonin 60; D047029:Lactams, Macrocyclic; D024101:Mitochondrial Proteins; D011498:Protein Precursors; D021382:Protein Sorting Signals; D015800:Protozoan Proteins; C001277:geldanamycin