Investigation on the interaction of human testis protein SPAG4L with KASH domain protein Nesprin-3 / 临床检验杂志

ABSTRACT

Objective To investigate the interaction of human testis sperm-associated antigen 4 like(SPAG4L) protein containing Sad,UNC-84(SUN) domain with nuclear envelop spectrin repeat proteins 3 (Nesprin-3) containing Klarsicht,ANC-1 and Syne homology (KASH) domain.Methods SPAC-4L protein domains were analyzed with the bioinformatics method.After the SPAG4L plasmid was transfected into Ntera-2 cells,the subcellular localization of SPAG4L was observed.The interaction of SPAG4L with Nesprin-3 was determined by immunofluorescence,co-immunoprecipitation(co-IP) and bimolecular fluorescence complementation (BiFC) technology,respectively.Results Bioinformatics analysis results showed that there was a transmembrane structure in SPAG4L protein.Subcellular localization results demonstrated that SPAG4L protein located in the nuclear membrane and cytoplasm.Immunofluorescence,Co-IP,and BiFC results showed that there was an interaction between SPAG4L and Nesprin-3,and that a LINC(linkers of the nucleoskeleton to the cytoskeletan) complex was formed.Conclusion SPAG4L may interact with Nesprin-3 to form a LINC complex,which is important for understanding the function of SPAG4L protein in spermatogenesis.