Identification of the Elusive Mammalian Enzyme Phosphatidylcholine-Specific Phospholipase C
Technical Report,01 Jul 2013,30 Jun 2015
STATE UNIV OF NEW YORK AT STONY BROOK RESEARCH FOUNDATION Stony Brook United States
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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. Identification of a novel experimental condition High serum that induces activation of PC-PLC. Besides being a novel observation in of itself, it allows us to overcome an unexpected obstacle lack of activation following LPS stimulation and allowed us to proceed further with our proposed transcriptome studies. Most of PC-PLC activity in basal and stimulated condition high serum resides in the cytosol. This is an important result since it greatly facilitates alternative approaches for the isolation of the PC-PLC protein with affinity or traditional chromatographical purification and it also refines the structural features of the putative PC-PLC protein no or one transmembrane domain that will help in the identification of the candidate PC-PLC clones from the transcriptome analysis. Provided for the first time clear-cut evidence of a potential role of PC-PLC activity in regulation of TNF-mediated inflammatory signaling.