Phospholipases

A) The hypothesis being tested here is the enhancement in the lipase performance of phospholipase C-?1 via phosphorylation of tyrosine 783.B) To perform the experiment equal concentrations of purified phospholipase-C-?1 were lay on incubation with the active kinase domain of FGFR2 and ATP in bovine serum albumin containing buffer.The samples of this reactions were tested for two activities 1) for lipase exertion in the phospholipid vehicles indicated in the figure on left y axis. Secondly the phosphate internalization in phospholipase-C-?1 was studied, illustrated at right y axis of figure.This was performed to con the phosphorylation of tyrosine and auto inhibition of PLC-? isozymes, 775/783 of PLC-?1 were substitutes at the place of phenylalanine, they could be use individually or together, but in the experiment tyr783 is used individually.Phospholipase activity of resulting mutant after purification was quantified with active domain of FGFR2K (helps in phosphorylation and ac tivation of phospholipase). certain known moles of phosphates were added into purified PLC-?1 in wild type to a lower place above mentioned conditions and was observed that phospholipase activity was enhanced 10 times.The mutation of tyr783 all nullified the kinase stimulated acceleration of phospholipase activity along with reduction in FGFR2K-promoted phosphorylation of PLC-?1. Therefore, phosphorylation of Tyr783 is vital forrelief of auto-inhibition. C) Studies reveal that Tyr-783 was essential for auto inhibition.As discussed above, long-lasting phosphorylation of tyr-783 exit completely nullify the kinase stimulated and FGFR2K stimulated phosphorylation of PLC-?1. lipase activity of PLC-?1 leave be enhanced across its limits and over-expression of PLC-?1 can induce cancerous transformation.The results could be leading to production of carcinoma cells. It has been found in various studies that activity of PLC-?1 is more in cancerous cells as compared to normal cells. So, permanent phosphorylation tyr783 could be a way leading to malignant cancers.