Abstract
The Coenzyme A (CoA), as a cofactor involved in >100 metabolic reactions, is essential to the basic biochemistry of life. Here, we investigated the CoA biosynthetic pathway of Entamoeba histolytica (E. histolytica), an enteric protozoan parasite responsible for human amebiasis. We identified four key enzymes involved in the CoA pathway: pantothenate kinase (PanK, EC 2.7.1.33), bifunctional phosphopantothenate-cysteine ligase/decarboxylase (PPCS-PPCDC), phosphopantetheine adenylyltransferase (PPAT) and dephospho-CoA kinase (DPCK). Cytosolic enzyme PanK, was selected for further biochemical, genetic, and phylogenetic characterization. Since E. histolytica PanK (EhPanK) is physiologically important and sufficiently divergent from its human orthologs, this enzyme represents an attractive target for the development of novel anti-amebic chemotherapies. Epigenetic gene silencing of PanK resulted in a significant reduction of PanK activity, intracellular CoA concentrations, and growth retardation in vitro, reinforcing the importance of this gene in E. histolytica. Furthermore, we screened the Kitasato Natural Products Library for inhibitors of recombinant EhPanK, and identified 14 such compounds. One compound demonstrated moderate inhibition of PanK activity and cell growth at a low concentration, as well as differential toxicity towards E. histolytica and human cells.
| Original language | English |
|---|---|
| Pages (from-to) | 125-136 |
| Number of pages | 12 |
| Journal | International Journal for Parasitology: Drugs and Drug Resistance |
| Volume | 8 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - 2018 Apr 1 |
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Keywords
- Amebiasis
- Coenzyme A
- Drug development
- Entamoeba histolytica
- Gene silencing
- Pantothenate kinase
ASJC Scopus subject areas
- Parasitology
- Infectious Diseases
- Pharmacology (medical)
Cite this
Characterization and validation of Entamoeba histolytica pantothenate kinase as a novel anti-amebic drug target. / Nurkanto, Arif; Jeelani, Ghulam; Yamamoto, Takehiro; Naito, Yoshiko; Hishiki, Takako; Mori, Mihoko; Suematsu, Makoto; Shiomi, Kazuro; Hashimoto, Tetsuo; Nozaki, Tomoyoshi.
In: International Journal for Parasitology: Drugs and Drug Resistance, Vol. 8, No. 1, 01.04.2018, p. 125-136.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Characterization and validation of Entamoeba histolytica pantothenate kinase as a novel anti-amebic drug target
AU - Nurkanto, Arif
AU - Jeelani, Ghulam
AU - Yamamoto, Takehiro
AU - Naito, Yoshiko
AU - Hishiki, Takako
AU - Mori, Mihoko
AU - Suematsu, Makoto
AU - Shiomi, Kazuro
AU - Hashimoto, Tetsuo
AU - Nozaki, Tomoyoshi
PY - 2018/4/1
Y1 - 2018/4/1
N2 - The Coenzyme A (CoA), as a cofactor involved in >100 metabolic reactions, is essential to the basic biochemistry of life. Here, we investigated the CoA biosynthetic pathway of Entamoeba histolytica (E. histolytica), an enteric protozoan parasite responsible for human amebiasis. We identified four key enzymes involved in the CoA pathway: pantothenate kinase (PanK, EC 2.7.1.33), bifunctional phosphopantothenate-cysteine ligase/decarboxylase (PPCS-PPCDC), phosphopantetheine adenylyltransferase (PPAT) and dephospho-CoA kinase (DPCK). Cytosolic enzyme PanK, was selected for further biochemical, genetic, and phylogenetic characterization. Since E. histolytica PanK (EhPanK) is physiologically important and sufficiently divergent from its human orthologs, this enzyme represents an attractive target for the development of novel anti-amebic chemotherapies. Epigenetic gene silencing of PanK resulted in a significant reduction of PanK activity, intracellular CoA concentrations, and growth retardation in vitro, reinforcing the importance of this gene in E. histolytica. Furthermore, we screened the Kitasato Natural Products Library for inhibitors of recombinant EhPanK, and identified 14 such compounds. One compound demonstrated moderate inhibition of PanK activity and cell growth at a low concentration, as well as differential toxicity towards E. histolytica and human cells.
AB - The Coenzyme A (CoA), as a cofactor involved in >100 metabolic reactions, is essential to the basic biochemistry of life. Here, we investigated the CoA biosynthetic pathway of Entamoeba histolytica (E. histolytica), an enteric protozoan parasite responsible for human amebiasis. We identified four key enzymes involved in the CoA pathway: pantothenate kinase (PanK, EC 2.7.1.33), bifunctional phosphopantothenate-cysteine ligase/decarboxylase (PPCS-PPCDC), phosphopantetheine adenylyltransferase (PPAT) and dephospho-CoA kinase (DPCK). Cytosolic enzyme PanK, was selected for further biochemical, genetic, and phylogenetic characterization. Since E. histolytica PanK (EhPanK) is physiologically important and sufficiently divergent from its human orthologs, this enzyme represents an attractive target for the development of novel anti-amebic chemotherapies. Epigenetic gene silencing of PanK resulted in a significant reduction of PanK activity, intracellular CoA concentrations, and growth retardation in vitro, reinforcing the importance of this gene in E. histolytica. Furthermore, we screened the Kitasato Natural Products Library for inhibitors of recombinant EhPanK, and identified 14 such compounds. One compound demonstrated moderate inhibition of PanK activity and cell growth at a low concentration, as well as differential toxicity towards E. histolytica and human cells.
KW - Amebiasis
KW - Coenzyme A
KW - Drug development
KW - Entamoeba histolytica
KW - Gene silencing
KW - Pantothenate kinase
UR - http://www.scopus.com/inward/record.url?scp=85042869527&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85042869527&partnerID=8YFLogxK
U2 - 10.1016/j.ijpddr.2018.02.004
DO - 10.1016/j.ijpddr.2018.02.004
M3 - Article
C2 - 29518650
AN - SCOPUS:85042869527
VL - 8
SP - 125
EP - 136
JO - International Journal for Parasitology: Drugs and Drug Resistance
JF - International Journal for Parasitology: Drugs and Drug Resistance
SN - 2211-3207
IS - 1
ER -