The Relationship Between α4Integrin And Adaptor LPXN And The Effect Of LPXN On Cell Adhesion

ObjectiveAs a kind of transmembrane protein recepter, integrin regulates adheresion between celland cell, cell and extracellular matrix (ECM). It can also affects cell biological behavior asa cellular signal transduction receptor. Integrin include α subunit and β subunit, and4αintegrin mediates the cell-ECM specific adhension and the other biological fuction. LPXNis a member of paxillin family which involved in cell damage repair, it regulates cell focaladhesion, cell adhesion and cell mobile and so on.According to the research, we explore the α4integrin combined with LPXN and JurkatE6cell adhension induced by LPXN in order to further study the role of α4integrin inimpacting the function of LPXN.Methods(1) To investigate the binding of α4integrin and LPXN in vitro, GST-LPXN purificatedprotein and His-α4integrin protein are used to go on protein binding experiment. Toobserve the combination of α4integrin and endogenous LPXN of JurkatE6whichstimulated with FN by IP experiments.Construct and identify the recombinant plasmidpcDNA3.1-alpha4integrin, which transfect cos-7cell together with GFP-LPXN, thendetect the expression and binding of two proteins.(2) Transfect Jurkat E6cell with GFP-LPXN and mCherry-LPXN shRNA stimulated withfibronectin (FN). Statistics each adhesion rate by fluorescence microscopy.Results(1) pcDNA3.1-alpha4integrin recombinant plasmid was constructed and specific expressed successfully. Co-transfection experiments and protein binding assays in vitroshowed that α4integrin binded to LPXN. Endogenous LPXN of JurkatE6cell combinatedto α4integrin stimulated with FN.(2) Overexpress LPXN enhanced the adhesion of Jurkat E6cell stimulated by FN; the celladhesion was weakened after silenced LPXN stimulated by FN.Conclusionα4integrin can be combined with LPXN which enhanced JurkatE6cell adhesionstimulated by FN. We are going to explore the role of α4integrin in impacting the functionof LPXN in follow-up experiments, such as phosphorylation of LPXN and the LPXNmediated cell adhesion and migration.