Neospora caninum: Tachyzoites express a potent type-I nucleoside triphosphate hydrolase, but lack nucleoside diphosphate hydrolase activity

Takashi Asai, Daniel K. Howe, Kyoko Nakajima, Tomoyoshi Nozaki, Tsutomu Takeuchi, L. David Sibley

Research output: Contribution to journalArticle

39 Citations (Scopus)

Abstract

We have identified type I nucleoside triphosphate hydrolase (NTPase; EC 3.6.1.3) activity, previously thought to be restricted to the virulent strains of Toxoplasma gondii, in the cell extracts of Neospora caninum tachyzoites. Sequence analysis of a complete cDNA from Nc-1 strain indicated that N. caninum NTPases shared approximately 69% identity to the NTPases of T. gondii and are most similar to the NTPase-I isozyme. Southern blot analysis of genomic DNA and sequence analysis of two independent NTP clones from the Nc-1 strain revealed the presence of multiple genes, at least two of which are transcribed. Substrate specificity and K(m) values for MgATP2 and MgADP- hydrolysis for recombinant or partially purified native NcNTPase were the same as those for the type I isozyme (NTPase-I). Significantly, no type II enzyme (NTPase-II) activity for NDP hydrolysis was detected in cell extracts of N. caninum, although it is universally present in all T. gondii strains that have been tested. This intriguing difference between these two closely related apicomplexan parasites may provide insight into the function of the NTPases during intracellular parasitism.

Original languageEnglish
Pages (from-to)277-285
Number of pages9
JournalExperimental Parasitology
Volume90
Issue number3
DOIs
Publication statusPublished - 1998 Nov

Fingerprint

N-Glycosyl Hydrolases
Nucleoside-Triphosphatase
Neospora
Diphosphates
Toxoplasma
Cell Extracts
Isoenzymes
Hydrolysis
Substrate Specificity
Southern Blotting
DNA Sequence Analysis
Adenosine Diphosphate
Sequence Analysis
Parasites
Complementary DNA
Clone Cells
triphosphoric acid
Enzymes
Genes

Keywords

  • Apicomplexan protozoa
  • ATPase
  • Neosporosis
  • Purine salvage
  • Toxoplasma gondii

ASJC Scopus subject areas

  • Immunology
  • Parasitology
  • Infectious Diseases

Cite this

Neospora caninum : Tachyzoites express a potent type-I nucleoside triphosphate hydrolase, but lack nucleoside diphosphate hydrolase activity. / Asai, Takashi; Howe, Daniel K.; Nakajima, Kyoko; Nozaki, Tomoyoshi; Takeuchi, Tsutomu; Sibley, L. David.

In: Experimental Parasitology, Vol. 90, No. 3, 11.1998, p. 277-285.

Research output: Contribution to journalArticle

Asai, Takashi ; Howe, Daniel K. ; Nakajima, Kyoko ; Nozaki, Tomoyoshi ; Takeuchi, Tsutomu ; Sibley, L. David. / Neospora caninum : Tachyzoites express a potent type-I nucleoside triphosphate hydrolase, but lack nucleoside diphosphate hydrolase activity. In: Experimental Parasitology. 1998 ; Vol. 90, No. 3. pp. 277-285.
@article{d81a4d54ec774d7cbcc44c6189962ee8,
title = "Neospora caninum: Tachyzoites express a potent type-I nucleoside triphosphate hydrolase, but lack nucleoside diphosphate hydrolase activity",
abstract = "We have identified type I nucleoside triphosphate hydrolase (NTPase; EC 3.6.1.3) activity, previously thought to be restricted to the virulent strains of Toxoplasma gondii, in the cell extracts of Neospora caninum tachyzoites. Sequence analysis of a complete cDNA from Nc-1 strain indicated that N. caninum NTPases shared approximately 69{\%} identity to the NTPases of T. gondii and are most similar to the NTPase-I isozyme. Southern blot analysis of genomic DNA and sequence analysis of two independent NTP clones from the Nc-1 strain revealed the presence of multiple genes, at least two of which are transcribed. Substrate specificity and K(m) values for MgATP2 and MgADP- hydrolysis for recombinant or partially purified native NcNTPase were the same as those for the type I isozyme (NTPase-I). Significantly, no type II enzyme (NTPase-II) activity for NDP hydrolysis was detected in cell extracts of N. caninum, although it is universally present in all T. gondii strains that have been tested. This intriguing difference between these two closely related apicomplexan parasites may provide insight into the function of the NTPases during intracellular parasitism.",
keywords = "Apicomplexan protozoa, ATPase, Neosporosis, Purine salvage, Toxoplasma gondii",
author = "Takashi Asai and Howe, {Daniel K.} and Kyoko Nakajima and Tomoyoshi Nozaki and Tsutomu Takeuchi and Sibley, {L. David}",
year = "1998",
month = "11",
doi = "10.1006/expr.1998.4346",
language = "English",
volume = "90",
pages = "277--285",
journal = "Experimental Parasitology",
issn = "0014-4894",
publisher = "Academic Press Inc.",
number = "3",

}

TY - JOUR

T1 - Neospora caninum

T2 - Tachyzoites express a potent type-I nucleoside triphosphate hydrolase, but lack nucleoside diphosphate hydrolase activity

AU - Asai, Takashi

AU - Howe, Daniel K.

AU - Nakajima, Kyoko

AU - Nozaki, Tomoyoshi

AU - Takeuchi, Tsutomu

AU - Sibley, L. David

PY - 1998/11

Y1 - 1998/11

N2 - We have identified type I nucleoside triphosphate hydrolase (NTPase; EC 3.6.1.3) activity, previously thought to be restricted to the virulent strains of Toxoplasma gondii, in the cell extracts of Neospora caninum tachyzoites. Sequence analysis of a complete cDNA from Nc-1 strain indicated that N. caninum NTPases shared approximately 69% identity to the NTPases of T. gondii and are most similar to the NTPase-I isozyme. Southern blot analysis of genomic DNA and sequence analysis of two independent NTP clones from the Nc-1 strain revealed the presence of multiple genes, at least two of which are transcribed. Substrate specificity and K(m) values for MgATP2 and MgADP- hydrolysis for recombinant or partially purified native NcNTPase were the same as those for the type I isozyme (NTPase-I). Significantly, no type II enzyme (NTPase-II) activity for NDP hydrolysis was detected in cell extracts of N. caninum, although it is universally present in all T. gondii strains that have been tested. This intriguing difference between these two closely related apicomplexan parasites may provide insight into the function of the NTPases during intracellular parasitism.

AB - We have identified type I nucleoside triphosphate hydrolase (NTPase; EC 3.6.1.3) activity, previously thought to be restricted to the virulent strains of Toxoplasma gondii, in the cell extracts of Neospora caninum tachyzoites. Sequence analysis of a complete cDNA from Nc-1 strain indicated that N. caninum NTPases shared approximately 69% identity to the NTPases of T. gondii and are most similar to the NTPase-I isozyme. Southern blot analysis of genomic DNA and sequence analysis of two independent NTP clones from the Nc-1 strain revealed the presence of multiple genes, at least two of which are transcribed. Substrate specificity and K(m) values for MgATP2 and MgADP- hydrolysis for recombinant or partially purified native NcNTPase were the same as those for the type I isozyme (NTPase-I). Significantly, no type II enzyme (NTPase-II) activity for NDP hydrolysis was detected in cell extracts of N. caninum, although it is universally present in all T. gondii strains that have been tested. This intriguing difference between these two closely related apicomplexan parasites may provide insight into the function of the NTPases during intracellular parasitism.

KW - Apicomplexan protozoa

KW - ATPase

KW - Neosporosis

KW - Purine salvage

KW - Toxoplasma gondii

UR - http://www.scopus.com/inward/record.url?scp=0032212406&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0032212406&partnerID=8YFLogxK

U2 - 10.1006/expr.1998.4346

DO - 10.1006/expr.1998.4346

M3 - Article

C2 - 9806873

AN - SCOPUS:0032212406

VL - 90

SP - 277

EP - 285

JO - Experimental Parasitology

JF - Experimental Parasitology

SN - 0014-4894

IS - 3

ER -