Identification of the Elusive Mammalian Enzyme Phosphatidylcholine-Specific Phospholipase C
Annual rept. 1 Jul 2013-10 Jun 2014
STATE UNIV OF NEW YORK AT STONY BROOK RESEARCH FOUNDATION
Pagination or Media Count:
The long-term purpose of the present proposal aims at establishing the role of the elusive mammalian protein, phosphatidycholine-specific phospholipase C PC-PLC in the inflammatory processes involved in progression of rheumatoid arthritis RA. Thus, the main scopes of this proposal are 1. to identify the PC-PLC gene and protein and 2. to test PC-PLC involvement in production of TNF alpha by monocytes. Major findings. In the first year of funding, we have mostly worked toward the identification of PC-PLC Aim 1. Based on results published by other groups, we proposed to compare transcriptome analysis between two PC-PLC activating conditions treatment with LPS and oxidized LDLs to identify candidate PC-PLC mRNAs. While we found that the published activation of human PC-PLC by LPS in HUVEC cells was due to Phospholipase D activity and not PC-PLC, we identified another condition high serum that leads to reproducible and specific activation of PC-PLC and that can be substituted to LPS in the transcriptome analysis. Moreover, we found that most of activated PC-PLC is in the cytosol with important implications for both the analysis of elevated transcripts and the possibility of chromatographic purification of PC-PLC.
- Genetic Engineering and Molecular Biology
- Medicine and Medical Research