title: Metabolic reconfiguration of the central glucose metabolism: a crucial strategy of Leishmania donovani for its survival during oxidative stress.
abstract: Understanding the mechanism that allows the intracellular protozoan parasite Leishmania donovani (Ld) to respond to reactive oxygen species (ROS) is of increasing therapeutic importance because of the continuing resistance toward antileishmanial drugs and for determining the illusive survival strategy of these parasites. A shift in primary carbon metabolism is the fastest response to oxidative stress. A (14)CO2 evolution study, expression of glucose transporters together with consumption assays, indicated a shift in metabolic flux of the parasites from glycolysis toward pentose phosphate pathway (PPP) when exposed to different oxidants in vitro/ex vivo. Changes in gene expression, protein levels, and enzyme activities all pointed to a metabolic reconfiguration of the central glucose metabolism in response to oxidants. Generation of glucose-6-phosphate dehydrogenase (G6PDH) (∼5-fold) and transaldolase (TAL) (∼4.2-fold) overexpressing Ld cells reaffirmed that lethal doses of ROS were counterbalanced by effective manipulation of NADPH:NADP(+) ratio and stringent maintenance of reduced thiol content. The extent of protein carbonylation and accumulation of lipid peroxidized products were also found to be less in overexpressed cell lines. Interestingly, the LD50 of sodium antimony gluconate (SAG), amphotericin-B (AmB), and miltefosine were significantly high toward overexpressing parasites. Consequently, this study illustrates that Ld strategizes a metabolic reconfiguration for replenishment of NADPH pool to encounter oxidative challenges.
authors.affiliation: *Division of Molecular Biology, Rajendra Memorial Research Institute of Medical Sciences (Indian Council of Medical Research), Agamkuan, Patna, Bihar, India; Department of Biotechnology, National Institute of Pharmaceutical Education and Research, Export Promotion Industrial Park, Hajipur, Vaishali, Bihar, India; Department of Microbiology, All India Institute of Medical Sciences, Phulwarisharif, Patna, Bihar, India; and Division of Infectious Diseases and Immunology, CSIR-Indian Institute of Chemical Biology, Jadavpur, Kolkata, India.
authors.forename: Ayan K
authors.initials: AK
authors.lastname: Ghosh
authors.affiliation: *Division of Molecular Biology, Rajendra Memorial Research Institute of Medical Sciences (Indian Council of Medical Research), Agamkuan, Patna, Bihar, India; Department of Biotechnology, National Institute of Pharmaceutical Education and Research, Export Promotion Industrial Park, Hajipur, Vaishali, Bihar, India; Department of Microbiology, All India Institute of Medical Sciences, Phulwarisharif, Patna, Bihar, India; and Division of Infectious Diseases and Immunology, CSIR-Indian Institute of Chemical Biology, Jadavpur, Kolkata, India.
authors.forename: Abul H
authors.initials: AH
authors.lastname: Sardar
authors.affiliation: *Division of Molecular Biology, Rajendra Memorial Research Institute of Medical Sciences (Indian Council of Medical Research), Agamkuan, Patna, Bihar, India; Department of Biotechnology, National Institute of Pharmaceutical Education and Research, Export Promotion Industrial Park, Hajipur, Vaishali, Bihar, India; Department of Microbiology, All India Institute of Medical Sciences, Phulwarisharif, Patna, Bihar, India; and Division of Infectious Diseases and Immunology, CSIR-Indian Institute of Chemical Biology, Jadavpur, Kolkata, India.
authors.forename: Abhishek
authors.initials: A
authors.lastname: Mandal
authors.affiliation: *Division of Molecular Biology, Rajendra Memorial Research Institute of Medical Sciences (Indian Council of Medical Research), Agamkuan, Patna, Bihar, India; Department of Biotechnology, National Institute of Pharmaceutical Education and Research, Export Promotion Industrial Park, Hajipur, Vaishali, Bihar, India; Department of Microbiology, All India Institute of Medical Sciences, Phulwarisharif, Patna, Bihar, India; and Division of Infectious Diseases and Immunology, CSIR-Indian Institute of Chemical Biology, Jadavpur, Kolkata, India.
authors.forename: Savita
authors.initials: S
authors.lastname: Saini
authors.affiliation: *Division of Molecular Biology, Rajendra Memorial Research Institute of Medical Sciences (Indian Council of Medical Research), Agamkuan, Patna, Bihar, India; Department of Biotechnology, National Institute of Pharmaceutical Education and Research, Export Promotion Industrial Park, Hajipur, Vaishali, Bihar, India; Department of Microbiology, All India Institute of Medical Sciences, Phulwarisharif, Patna, Bihar, India; and Division of Infectious Diseases and Immunology, CSIR-Indian Institute of Chemical Biology, Jadavpur, Kolkata, India.
authors.forename: Kumar
authors.initials: K
authors.lastname: Abhishek
authors.affiliation: *Division of Molecular Biology, Rajendra Memorial Research Institute of Medical Sciences (Indian Council of Medical Research), Agamkuan, Patna, Bihar, India; Department of Biotechnology, National Institute of Pharmaceutical Education and Research, Export Promotion Industrial Park, Hajipur, Vaishali, Bihar, India; Department of Microbiology, All India Institute of Medical Sciences, Phulwarisharif, Patna, Bihar, India; and Division of Infectious Diseases and Immunology, CSIR-Indian Institute of Chemical Biology, Jadavpur, Kolkata, India.
authors.forename: Ashish
authors.initials: A
authors.lastname: Kumar
authors.affiliation: *Division of Molecular Biology, Rajendra Memorial Research Institute of Medical Sciences (Indian Council of Medical Research), Agamkuan, Patna, Bihar, India; Department of Biotechnology, National Institute of Pharmaceutical Education and Research, Export Promotion Industrial Park, Hajipur, Vaishali, Bihar, India; Department of Microbiology, All India Institute of Medical Sciences, Phulwarisharif, Patna, Bihar, India; and Division of Infectious Diseases and Immunology, CSIR-Indian Institute of Chemical Biology, Jadavpur, Kolkata, India.
authors.forename: Bidyut
authors.initials: B
authors.lastname: Purkait
authors.affiliation: *Division of Molecular Biology, Rajendra Memorial Research Institute of Medical Sciences (Indian Council of Medical Research), Agamkuan, Patna, Bihar, India; Department of Biotechnology, National Institute of Pharmaceutical Education and Research, Export Promotion Industrial Park, Hajipur, Vaishali, Bihar, India; Department of Microbiology, All India Institute of Medical Sciences, Phulwarisharif, Patna, Bihar, India; and Division of Infectious Diseases and Immunology, CSIR-Indian Institute of Chemical Biology, Jadavpur, Kolkata, India.
authors.forename: Ruby
authors.initials: R
authors.lastname: Singh
authors.affiliation: *Division of Molecular Biology, Rajendra Memorial Research Institute of Medical Sciences (Indian Council of Medical Research), Agamkuan, Patna, Bihar, India; Department of Biotechnology, National Institute of Pharmaceutical Education and Research, Export Promotion Industrial Park, Hajipur, Vaishali, Bihar, India; Department of Microbiology, All India Institute of Medical Sciences, Phulwarisharif, Patna, Bihar, India; and Division of Infectious Diseases and Immunology, CSIR-Indian Institute of Chemical Biology, Jadavpur, Kolkata, India.
authors.forename: Sushmita
authors.initials: S
authors.lastname: Das
authors.affiliation: *Division of Molecular Biology, Rajendra Memorial Research Institute of Medical Sciences (Indian Council of Medical Research), Agamkuan, Patna, Bihar, India; Department of Biotechnology, National Institute of Pharmaceutical Education and Research, Export Promotion Industrial Park, Hajipur, Vaishali, Bihar, India; Department of Microbiology, All India Institute of Medical Sciences, Phulwarisharif, Patna, Bihar, India; and Division of Infectious Diseases and Immunology, CSIR-Indian Institute of Chemical Biology, Jadavpur, Kolkata, India.
authors.forename: Rupkatha
authors.initials: R
authors.lastname: Mukhopadhyay
authors.affiliation: *Division of Molecular Biology, Rajendra Memorial Research Institute of Medical Sciences (Indian Council of Medical Research), Agamkuan, Patna, Bihar, India; Department of Biotechnology, National Institute of Pharmaceutical Education and Research, Export Promotion Industrial Park, Hajipur, Vaishali, Bihar, India; Department of Microbiology, All India Institute of Medical Sciences, Phulwarisharif, Patna, Bihar, India; and Division of Infectious Diseases and Immunology, CSIR-Indian Institute of Chemical Biology, Jadavpur, Kolkata, India.
authors.forename: Syamal
authors.initials: S
authors.lastname: Roy
authors.affiliation: *Division of Molecular Biology, Rajendra Memorial Research Institute of Medical Sciences (Indian Council of Medical Research), Agamkuan, Patna, Bihar, India; Department of Biotechnology, National Institute of Pharmaceutical Education and Research, Export Promotion Industrial Park, Hajipur, Vaishali, Bihar, India; Department of Microbiology, All India Institute of Medical Sciences, Phulwarisharif, Patna, Bihar, India; and Division of Infectious Diseases and Immunology, CSIR-Indian Institute of Chemical Biology, Jadavpur, Kolkata, India drpradeep.das@gmail.com.
authors.forename: Pradeep
authors.initials: P
authors.lastname: Das
mesh_terms: D000977:Antiparasitic Agents / Q000494:pharmacology; D015153:Blotting, Western; D002478:Cells, Cultured; D004351:Drug Resistance; D005947:Glucose / Q000378:metabolism*; D005954:Glucosephosphate Dehydrogenase / Q000378:metabolism; D006019:Glycolysis / Q000187:drug effects / Q000502:physiology*; D006801:Humans; D007893:Leishmania donovani / Q000472:pathogenicity; D007898:Leishmaniasis, Visceral / Q000378:metabolism* / Q000401:mortality / Q000469:parasitology; D015227:Lipid Peroxidation / Q000187:drug effects; D009249:NADP / Q000378:metabolism; D016877:Oxidants / Q000494:pharmacology*; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D010427:Pentose Phosphate Pathway; D050050:Protein Carbonylation / Q000187:drug effects; D012333:RNA, Messenger / Q000235:genetics; D017382:Reactive Oxygen Species / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D000977:Antiparasitic Agents; D016877:Oxidants; D012333:RNA, Messenger; D017382:Reactive Oxygen Species; D009249:NADP; D005954:Glucosephosphate Dehydrogenase; D005947:Glucose
keywords: NADPH; PPP; metabolic flux