Thymidine kinase and uridine-cytidine kinase from Entamoeba histolytica

Cloning, characterization, and search for specific inhibitors

A. Lossani, A. Torti, S. Gatti, A. Bruno, R. Maserati, G. E. Wright, F. Focher

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Entamoeba histolytica is an intestinal parasite and the causative agent of amoebiasis, which is a significant source of morbidity and mortality in developing countries. Although anti-amoebic drugs such as metronidazole, emetine, chloroquine and nitazoxanide are generally effective, there is always potential for development of drug resistance. In order to find novel targets to control E. histolytica proliferation we cloned, expressed and purified thymidine kinase (Eh-TK) and uridine-cytidine kinase (Eh-UCK) from E. histolytica. Eh-TK phosphorylates thymidine with a Km of 0.27 μm, whereas Eh-UCK phosphorylates uridine and cytidine with Km of 0.74 and 0.22 mm, respectively. For both enzymes, ATP acts as specific phosphate donor. In order to find alternative treatments of E. histolytica infection we tested numerous nucleoside analogues and related compounds as inhibitors and/or substrates of Eh-TK and Eh-UCK, and active compounds against E. histolytica in cell culture. Our results indicate that inhibitors or alternative substrates of the enzymes, although partially reducing protozoan proliferation, are reversible and not likely to become drugs against E. histolytica infections.

Original languageEnglish
Pages (from-to)595-602
Number of pages8
JournalParasitology
Volume136
Issue number6
DOIs
Publication statusPublished - May 2009

Fingerprint

Uridine Kinase
Entamoeba histolytica
thymidine kinase
Thymidine Kinase
Organism Cloning
nitazoxanide
emetine
amebiasis
Emetine
cytidine
chloroquine
Cytidine
drugs
metronidazole
Amebiasis
uridine
enzyme substrates
nucleosides
Uridine
Metronidazole

Keywords

  • Amoebiasis
  • Drugs
  • Entamoeba
  • Inhibitors
  • Kinases

ASJC Scopus subject areas

  • Parasitology
  • Animal Science and Zoology
  • Infectious Diseases

Cite this

Thymidine kinase and uridine-cytidine kinase from Entamoeba histolytica : Cloning, characterization, and search for specific inhibitors. / Lossani, A.; Torti, A.; Gatti, S.; Bruno, A.; Maserati, R.; Wright, G. E.; Focher, F.

In: Parasitology, Vol. 136, No. 6, 05.2009, p. 595-602.

Research output: Contribution to journalArticle

@article{8d87c5948f4e4be1a24a5e1750f840b2,
title = "Thymidine kinase and uridine-cytidine kinase from Entamoeba histolytica: Cloning, characterization, and search for specific inhibitors",
abstract = "Entamoeba histolytica is an intestinal parasite and the causative agent of amoebiasis, which is a significant source of morbidity and mortality in developing countries. Although anti-amoebic drugs such as metronidazole, emetine, chloroquine and nitazoxanide are generally effective, there is always potential for development of drug resistance. In order to find novel targets to control E. histolytica proliferation we cloned, expressed and purified thymidine kinase (Eh-TK) and uridine-cytidine kinase (Eh-UCK) from E. histolytica. Eh-TK phosphorylates thymidine with a Km of 0.27 μm, whereas Eh-UCK phosphorylates uridine and cytidine with Km of 0.74 and 0.22 mm, respectively. For both enzymes, ATP acts as specific phosphate donor. In order to find alternative treatments of E. histolytica infection we tested numerous nucleoside analogues and related compounds as inhibitors and/or substrates of Eh-TK and Eh-UCK, and active compounds against E. histolytica in cell culture. Our results indicate that inhibitors or alternative substrates of the enzymes, although partially reducing protozoan proliferation, are reversible and not likely to become drugs against E. histolytica infections.",
keywords = "Amoebiasis, Drugs, Entamoeba, Inhibitors, Kinases",
author = "A. Lossani and A. Torti and S. Gatti and A. Bruno and R. Maserati and Wright, {G. E.} and F. Focher",
year = "2009",
month = "5",
doi = "10.1017/S0031182009005964",
language = "English",
volume = "136",
pages = "595--602",
journal = "Parasitology",
issn = "0031-1820",
publisher = "Cambridge University Press",
number = "6",

}

TY - JOUR

T1 - Thymidine kinase and uridine-cytidine kinase from Entamoeba histolytica

T2 - Cloning, characterization, and search for specific inhibitors

AU - Lossani, A.

AU - Torti, A.

AU - Gatti, S.

AU - Bruno, A.

AU - Maserati, R.

AU - Wright, G. E.

AU - Focher, F.

PY - 2009/5

Y1 - 2009/5

N2 - Entamoeba histolytica is an intestinal parasite and the causative agent of amoebiasis, which is a significant source of morbidity and mortality in developing countries. Although anti-amoebic drugs such as metronidazole, emetine, chloroquine and nitazoxanide are generally effective, there is always potential for development of drug resistance. In order to find novel targets to control E. histolytica proliferation we cloned, expressed and purified thymidine kinase (Eh-TK) and uridine-cytidine kinase (Eh-UCK) from E. histolytica. Eh-TK phosphorylates thymidine with a Km of 0.27 μm, whereas Eh-UCK phosphorylates uridine and cytidine with Km of 0.74 and 0.22 mm, respectively. For both enzymes, ATP acts as specific phosphate donor. In order to find alternative treatments of E. histolytica infection we tested numerous nucleoside analogues and related compounds as inhibitors and/or substrates of Eh-TK and Eh-UCK, and active compounds against E. histolytica in cell culture. Our results indicate that inhibitors or alternative substrates of the enzymes, although partially reducing protozoan proliferation, are reversible and not likely to become drugs against E. histolytica infections.

AB - Entamoeba histolytica is an intestinal parasite and the causative agent of amoebiasis, which is a significant source of morbidity and mortality in developing countries. Although anti-amoebic drugs such as metronidazole, emetine, chloroquine and nitazoxanide are generally effective, there is always potential for development of drug resistance. In order to find novel targets to control E. histolytica proliferation we cloned, expressed and purified thymidine kinase (Eh-TK) and uridine-cytidine kinase (Eh-UCK) from E. histolytica. Eh-TK phosphorylates thymidine with a Km of 0.27 μm, whereas Eh-UCK phosphorylates uridine and cytidine with Km of 0.74 and 0.22 mm, respectively. For both enzymes, ATP acts as specific phosphate donor. In order to find alternative treatments of E. histolytica infection we tested numerous nucleoside analogues and related compounds as inhibitors and/or substrates of Eh-TK and Eh-UCK, and active compounds against E. histolytica in cell culture. Our results indicate that inhibitors or alternative substrates of the enzymes, although partially reducing protozoan proliferation, are reversible and not likely to become drugs against E. histolytica infections.

KW - Amoebiasis

KW - Drugs

KW - Entamoeba

KW - Inhibitors

KW - Kinases

UR - http://www.scopus.com/inward/record.url?scp=67651165418&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=67651165418&partnerID=8YFLogxK

U2 - 10.1017/S0031182009005964

DO - 10.1017/S0031182009005964

M3 - Article

VL - 136

SP - 595

EP - 602

JO - Parasitology

JF - Parasitology

SN - 0031-1820

IS - 6

ER -