期刊
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
卷 114, 期 11, 页码 E2068-E2076出版社
NATL ACAD SCIENCES
DOI: 10.1073/pnas.1615195114
关键词
heme; heme sensor; genetically encoded biosensor; malaria; Plasmodium falciparum
资金
- National Institute of Environmental Health Sciences Center [P30-ES002109]
- National Institute of General Medical Sciences (NIGMS) Biotechnology Training [5-T32-GM08334]
- NIH Director's New Innovator Award [1DP2OD007124]
- NIGMS Center for Integrative Synthetic Biology [P50-GM098792]
- NIH [5-P41-EB015871-27, DP3-DK101024 01, 1-U01-NS090438- 01, 1-R01-EY017656-0,6A1, 1-R01-HL121386-01A1]
- Singapore-MIT Alliance 2
- BioSystems and Micromechanics Interdisciplinary Research Group of Singapore-MIT Alliance Research and Technology Center
- Koch Institute for Integrative Cancer Research Bridge Initiative
- Wellcome Trust MIT Postdoctoral Research
- Wellcome Trust
- MIT SkolTech Initiative
- Ferry Award
- MIT startup funds
Heme is ubiquitous, yet relatively little is known about the maintenance of labile pools of this cofactor, which likely ensures its timely bioavailability for proper cellular function. Quantitative analysis of labile heme is of fundamental importance to understanding how nature preserves access to the diverse chemistry heme enables, while minimizing cellular damage caused by its redox activity. Here, we have developed and characterized a protein-based sensor that undergoes fluorescence quenching upon heme binding. By genetically encoding this sensor in the human malarial parasite, Plasmodium falciparum, we have quantified cytosolic labile heme levels in intact, blood-stage parasites. Our findings indicate that a labile heme pool (1.6 mu M) is stably maintained throughout parasite development within red blood cells, even during a period coincident with extensive hemoglobin degradation by the parasite. We also find that the heme-binding antimalarial drug chloroquine specifically increases labile cytosolic heme, indicative of dysregulation of this homeostatic pool that may be a relevant component of the antimalarial activity of this compound class. We propose that use of this technology under various environmental perturbations in P. falciparum can yield quantitative insights into fundamental heme biology.
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