Abstract
The phosphoinositide (PI)-specific phospholipase C gene (TcPI-PLC) of the protozoan parasite Trypanosoma cruzi was cloned, sequenced, expressed in Escherichia coli, and the protein product (TcPI-PLC) was shown to have enzymatic characteristics similar to those of mammalian delta-type PI-PLCs. The TcPI-PLC gene is expressed at high levels in the epimastigote and amastigote stages of the parasite, and its expression is induced during the differentiation of trypomastigotes into amastigotes, where TcPI-PLC associates with the plasma membrane and increases its catalytic activity. In contrast to other PI-PLCs described so far, the deduced amino acid sequence of TcPI-PLC revealed some unique features such as an N-myristoylation consensus sequence at its amino-terminal end, lack of an apparent pleckstrin homology domain and a highly charged linker region between the catalytic X and Y domains. TcPI-PLC is lipid modified in vivo, as demonstrated by metabolic labeling with [(3)H]myristate and [(3)H]palmitate and fatty acid analysis of the immunoprecipitated protein, and may constitute the first example of a new group of PI-PLCs.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Animals
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Base Sequence
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Calcium / pharmacology
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Cell Differentiation*
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Cell Membrane / enzymology
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Cloning, Molecular
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Enzyme Activation
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Fatty Acids / metabolism
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Fluorescent Antibody Technique
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Gene Expression Regulation, Developmental / genetics
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Hydrogen-Ion Concentration
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Inositol 1,4,5-Trisphosphate / metabolism
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Membrane Proteins / chemistry
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Membrane Proteins / genetics
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Molecular Sequence Data
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Myristic Acid / metabolism
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Phosphatidylinositol 4,5-Diphosphate / metabolism
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Phosphatidylinositol Diacylglycerol-Lyase
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Protozoan Proteins / metabolism
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RNA, Messenger / metabolism
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Recombinant Proteins / genetics
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Recombinant Proteins / isolation & purification
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Sequence Homology, Amino Acid
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Trypanosoma cruzi / enzymology*
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Trypanosoma cruzi / growth & development
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Type C Phospholipases / chemistry
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Type C Phospholipases / genetics
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Type C Phospholipases / metabolism*
Substances
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Fatty Acids
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Membrane Proteins
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Phosphatidylinositol 4,5-Diphosphate
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Protozoan Proteins
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RNA, Messenger
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Recombinant Proteins
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Myristic Acid
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Inositol 1,4,5-Trisphosphate
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Type C Phospholipases
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Phosphatidylinositol Diacylglycerol-Lyase
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Calcium