Preliminary Study On The Inflammatory Bodies Of GSDMD And NLRP3

Congenital immunity plays a very important role in the body of the organism.Innate immunity leads to inflammatory necrosis of cells.Cell death is an important way of cell necrosis and also an important way of programmed cell death.Therefore,exploring the proteins associated with the pathway of the death is important for the study of natural immunity.The current study is more clear about the GSHMD,also known as DFNA5.This protein contains a special Gasdemin domain,belonging to the Gasdemin family.The Gasdemin domain family includes GSDMA,GSDMB,GSDMC,DFNA5,DFNB59,etc.Related members.There are also reports that two independent research groups have used gene screening technology to discover a gelatin substrate protein,GSDMD,which acts as a substrate for the death of the substrate and a substrate for the downstream of the inflammatory response.GSDMD triggers cell coke.Death promotes cell necrosis.Studies have also shown that if the GSDMD protein is removed from the cells,the cells will not swell and will not rupture.At the same time,the occurrence of cell coke formation will promote a strong inflammatory response.According to this feature,GSDMD is also expressed in esophageal esophagus and skin.The expression site of GSDMD is generally at the site of pathogen invasion.Therefore,the GSDMD protein was screened.Therefore,we can choose to use GSDMD as a bait protein to do a screening process,and then screen out the relevant proteins and further verify the experiment from the function.After screening by yeast,the related genes N1,Z1,Z2,and S1 are basically genes involved in transcription or signal transduction,and may have a regulatory effect on focus.No further results were obtained when further validation by CO-IP experiments,and the interaction remains to be further verified.In the body,GSDMD is in an inactive dormant state.GSDMD needs to be cleaved and activated by caspase-1 inside the cell to form an active N-terminal domain.The active GSDMD-N end will bind to The cell membrane is perforated on the liposome and on the cell membrane,and finally causes the cell rupture content to flow out,causing the cell to swell.Studies have shown that NLRP3 inflammatory bodies can activate caspase-1,a very important inflammatory corpuscle inside the body.There are two activation models for NLRP3 inflammatory bodies,one of which includes NF-κB.The signaling pathway up-regulates the expression of inflammatory factors and IL-1βto regulate the activation of inflammatory corpuscles.The mechanism of action is that the caspase-1,ASC and NLRP3 molecules assemble to form active inflammatory bodies.Another regulation is the regulation of individual proteins in the signaling pathway.The NLRP3 inflammatory corpuscle activates the caspase-1,and the activation of the precursor caspase-1 triggers the maturation and secretion of IL-1β.Subsequently,we studied the interacting proteins of inflammatory corpuscle NLRP3 related to GSDMD activation,and screened proteins interacting with NLRP3 by mass spectrometry.By overexpressing plasmid NLRP3-FLAG in 293 T cells,the corresponding label was used.The antibody was incubated for CO-IP experiment.After the beads were added to the antibody for adsorption overnight,the nonspecifically bound protein was eluted with the corresponding eluent,and the proteinbearing beads were sent to the company for mass spectrometry.The result was relatively high.And a protein with a strong specific binding.From the results of mass spectrometry,the TMPRSS13 protein was obtained and the content was high.One of the proteins was called TMPRSS13.This protein is a type II transmembrane serine protease.We confirmed that NLRP3 interacts with TMPRSS13.And further explored its relationship with inflammatory response,it can regulate the expression of IL-1β,a gene downstream of the inflammatory response,and found that the protein has an inhibitory effect on the activation of inflammatory corpuscles.We explored NLRP3.At the binding site of the protease,it was found that TMPRSS13 interacts with each domain of NLRP3,but the binding in the LRR region is the strongest,and it is speculated that this site is a combination of TMPRSS13 and other ligands of the inflammatory corpuscle.Site.However,the specific mechanism of how the protein’s action site is regulated in the body requires further experiments to verify.