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Dimerization of phospholipase d isozymes.
1Howard Hughes Medical Institute, Vanderbilt University School of Medicine, Nashville, TN 37232, USA.
Biochemical and Biophysical Research Communications
|January 10, 2002
Summary
Mammalian phospholipase D (PLD) isozymes PLD1 and PLD2 can form homodimers and heterodimers. These PLD proteins associate in cells, suggesting a role for dimerization in their function.
Area of Science:
- Biochemistry
- Molecular Biology
- Cell Biology
Background:
- Two mammalian phospholipase D (PLD) isozymes, PLD1 and PLD2, have been identified.
- The potential for interaction and dimerization between these isozymes remains largely unexplored.
Purpose of the Study:
- To investigate the association between mammalian phospholipase D isozymes, PLD1 and PLD2.
- To determine if PLD1 and PLD2 can form homodimers and heterodimers.
Main Methods:
- Differential tagging of PLD isozymes with EGFP and Xpress peptide epitope.
- Coimmunoprecipitation assays to detect protein-protein interactions.
- Immunofluorescence staining to assess subcellular localization and colocalization.
Main Results:
- Overexpressed EGFP-rPLD1 coimmunoprecipitated with Xpress-rPLD1, independent of enzyme activity.
- Xpress-rPLD2 showed binding to EGFP-rPLD1, and EGFP-rPLD2 bound to Xpress-rPLD1, confirming heterodimerization.
- EGFP-rPLD2 also bound to Xpress-rPLD2, indicating homodimerization of PLD2.
- Immunofluorescence revealed colocalization of PLD isozymes in perinuclear and plasma membrane regions.
Conclusions:
- Mammalian PLD1 and PLD2 can form homodimers.
- Mammalian PLD1 and PLD2 can form heterodimers.
- These interactions occur within the cellular environment, suggesting functional relevance.