The Interaction Between KICbeta And 14-3-3epsilon

The interaction between proteins is the foundation of life.It is an important way that various cell function and activity accomplished.Our research used yeast two-hybrid screening system to find out more protein-protein interaction.Human fetal brain cDNA library was screened using 14-3-3 epsilon as the bait to discover proteins interacting with it.KICbeta is a member of mammalian kinesin super family protein.In neuronal cells,KICbeta bind synaptic vesicle precursors and transport them towards axon termini along the direction that microtubule synthesized.Previous research showed that KICbeta gene is loss of heterozygousin neuroblastoma.What is more,the haploinsufficiency.KICbeta gene could act as a tumorsuppressor.Overpressing KICbeta protein induces apoptotic death of cancer cell.We analyzed this gene by bio-information method,and amplificationed it from human fetal cDNA for subsequent research.To confirm the interaction between the KICbeta and 14-3-3 epsilon,we proformed GST pull-down and CoIP.The result demonstrated the two proteins are physics associated.KICbeta protein has a long peptide chain,so we divided it into three parts to see which part involved in this interaction.As a result,14-3-3epsilon binds the 600-1250 amino acids of KICbeta.Hela cells were co-transfection with GFP-KICbeta and RED-14-3-3epsilon,the sub-cellular localization showed they both distribute in cytoplasm.