| Cell surface membrane proteins bridge cellular inner world and surrounding circumstance together, which play an important role in cell physiological function. In order to find proteins which can interact with Protein 4.1 family and Protein MAGUK family member in nerve system, we used the cytoplasmic tail of red blood cell surface membrane protein Glycophorin C as a probe, cloned a novel human cDNA sequence and named it as NECL2 (Nectin Like Protein2), because the structure of the protein encoded by NECL2 is very similar with nectin family.NECL2 is widely expressed in multiple tissues, such as brain heart, liver lung and skeletal muscle by using Northern Blot. To explore the role of NECL2 in CNS (Central Nerve System), we analyzed expression pattern of NECL2 by using the ISH (In Situ Hybridization) and immunohistochemistry. NECL2 was detected in cerebral cortex hippocampus purkinje cells in cerebellum striatum pons nuclei and anterior horn of spinal cord. NECL2 distributed in cell body and protuberance of primary culture neuron. The rat adrenal gland tumor cell line PC12 can be induced into neuronal differentiation. Transfection and over-expression of NECL2 in PC12 induced the cell undergo neuronal differentiation. Protein 4.1 family is a group of cell skeleton associated proteins which is very important in CNS. NECL2 was identified to interact with Protein 4.1 family member DAL-1 by using protein binding assay in vitro and cell immunofluorescence co-localization, which provide clues to issue the mechanism of NECL2 induction of cell neuronal differentiation.NECL2 was also found to induce apoptosis of human acute leukemia cell line NB4. By yeast two-hybrid screening with the cytoplasmic region of NECL2, a possible interacting protein cFLIP was identified and confirmed to interact with NECL2 through the verification of protein binding in vitro and in vivo. cFLIP is a apoptosis associated protein, shares highly similarity with capase family and participates in...
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