Target proteins of the cytosolic thioredoxin in Plasmodium falciparum |
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| Authors: | Shin-ichiro Kawazu Hitoshi Takemae Kanako Komaki-Yasuda Shigeyuki Kano |
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| Affiliation: | 1. National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, 2-13 Inada-cho, Obihiro, Hokkaido 080-8555, Japan;2. Research Institute, International Medical Center of Japan, 1-21-1 Toyama, Shinjuku-ku, Tokyo 162-8655, Japan;3. Center for Advanced Proteomics Research and Department of Microbiology, Biochemistry & Molecular Genetics, Rutgers University-New Jersey Medical School Cancer Center, 205 S. Orange Ave., Newark, New Jersey 07103;5. Department of Cell Biology and Molecular Medicine, Rutgers University-New Jersey Medical School, 185 S. Orange Ave., Newark, New Jersey 07103;6. Albert Einstein College of Medicine, 1301 Morris Park Avenue, Bronx, New York 10461;1. Malaria Research, Infectious Diseases Unit, Department of Medicine Solna, Karolinska University Hospital/Karolinska Institutet, Retzius väg 10, 171 77 Stockholm, Sweden;2. Departamento de Parasitología, Escuela de Microbiología, Facultad de Ciencias, Universidad Nacional Autónoma de Honduras (UNAH), Tegucigalpa, Honduras;3. Department of Microbiology, Tumor and Cell Biology (MTC), Karolinska Institutet, Stockholm, Sweden;4. Life and Health Sciences Research Institute (ICVS), School of Health Sciences, University of Minho, Braga, Portugal;5. ICVS/3B’s – PT Government Associate Laboratory, Braga/Guimarães, Portugal;6. Global Health, Department of Public Health Sciences, Karolinska Institutet, Stockholm, Sweden;7. Malaria and Vector Research Group, Biotechnology Research Center, Pasteur Institute of Iran, Iran;8. Centro Internacional de Entrenamiento e Investigaciones Médicas, Cali, Colombia;9. Shoklo Malaria Research Unit, Mae Sot Tak, Thailand;10. Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand;11. Nuffield Department of Clinical Medicine, Centre for Clinical Vaccinology and Tropical Medicine, University of Oxford, Oxford, United Kingdom;12. School of Biological Sciences, Nanyang Technological University, Singapore;13. Department of Biomedical Sciences and Veterinary Public Health, Section of Virology, Swedish University of Agricultural Sciences, Uppsala, Sweden;1. Departamento de Bioquímica, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay;2. Center for Free Radical and Biomedical Research, Universidad de la República, Montevideo, Uruguay;3. Instituto de Química y Físicoquímica Biológicas “Prof. Alejandro C. Paladini” (IQUIFIB), Universidad de Buenos Aires and CONICET, Ciudad Autónoma de Buenos Aires, Argentina;4. Departamento de Química Inorgánica, Analítica y Química-Física and INQUIMAE-CONICET, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Buenos Aires, Argentina;5. Unidad de Biología Molecular-Institut Pasteur Montevideo, Montevideo, Uruguay |
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| Abstract: | The target proteins of a cytosolic Trx (PfTrx-1) in Plasmodium falciparum with Trx-affinity chromatography were examined. Based on the Trx protein reduction pathway, we generated a cysteine mutant of PfTrx-1, which captures the target protein as a mixed disulfide intermediate. A number of proteins were captured with PfTrx-1(C33S) immobilized on resin and were eluted by DTT treatment. The PfTrx-1(C33S) immobilized resin-captured proteins were trypsin-digested and analyzed on a liquid chromatography-mass spectrometry system. Analysis of the sequence data against databases assigned 20 proteins, four of which had been found previously in P. falciparum, with the remaining 16 being new targets. The potential Trx-target proteins included those in pathways such as the redox cycle, protein biosynthesis, energy metabolism and signal transduction. We captured 4 enzymes in the glycolysis pathway (hexokinase, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), phosphoglycerate mutase and l-lactate dehydrogenase (LDH)) as Trx-targets, and we found that PfTrx-1 enhanced the activity of PfGAPDH and PfLDH. |
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