The Toxoplasma gondii phosphoinositide-specific phospholipase C gene (TgPI-PLC) was cloned, sequenced and expressed in Escherichia coli and its enzymatic characteristics were investigated. TgPI-PLC is present in the genome as a single-copy gene consisting of 22 exons interrupted by 21 introns, and encodes a polypeptide of 1097 amino acids with a predicted molecular mass of 121 kDa. In addition to the conserved catalytic X and Y domains, TgPI-PLC contains an apparent N-terminal PH domain, an EF hand motif and a C-terminal C2 domain. When compared with mammalian δ-type PI-PLC, TgPI-PLC has an additional extended N-terminus and two insertions in the region between the X and Y domains, with a 31–35% identity over the whole sequence. Recombinant TgPI-PLC, as well as the native enzyme obtained from crude membrane extracts of the parasite, was more active with phosphatidylinositol than with phosphatidylinositol 4,5-bisphosphate as substrate. Indirect immunofluorescence analysis using an affinity-purified antibody against TgPI-PLC revealed that this enzyme localizes in the plasma membrane of the parasites.
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Research Article|
February 10 2006
A Toxoplasma gondii phosphoinositide phospholipase C (TgPI-PLC) with high affinity for phosphatidylinositol
Jianmin Fang;
Jianmin Fang
*Department of Cellular Biology, and Center for Tropical and Emerging Global Diseases, The University of Georgia, Athens, GA 30602, U.S.A.
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Norma Marchesini;
Norma Marchesini
†Laboratory of Molecular Parasitology, Department of Pathobiology and Center for Zoonoses Research, University of Illinois at Urbana-Champaign, Urbana, IL 61802, U.S.A.
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Silvia N. J. Moreno
Silvia N. J. Moreno
1
*Department of Cellular Biology, and Center for Tropical and Emerging Global Diseases, The University of Georgia, Athens, GA 30602, U.S.A.
†Laboratory of Molecular Parasitology, Department of Pathobiology and Center for Zoonoses Research, University of Illinois at Urbana-Champaign, Urbana, IL 61802, U.S.A.
1To whom correspondence should be addressed (email smoreno@uga.edu).
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Publisher: Portland Press Ltd
Received:
August 24 2005
Revision Received:
November 04 2005
Accepted:
November 16 2005
Accepted Manuscript online:
November 16 2005
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London
2006
Biochem J (2006) 394 (2): 417–425.
Article history
Received:
August 24 2005
Revision Received:
November 04 2005
Accepted:
November 16 2005
Accepted Manuscript online:
November 16 2005
Citation
Jianmin Fang, Norma Marchesini, Silvia N. J. Moreno; A Toxoplasma gondii phosphoinositide phospholipase C (TgPI-PLC) with high affinity for phosphatidylinositol. Biochem J 1 March 2006; 394 (2): 417–425. doi: https://doi.org/10.1042/BJ20051393
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