NKp30/NCR3 Overexpression Lysate

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

NKp30, also referred to as natural cytotoxicity receptor 3 (NCR3), is a type I transmembrane receptor belonging to the immunoglobulin (Ig) superfamily that is expressed on resting and active natural killer (NK) cells with a role in triggering a cytotoxic response against tumor cells (1). The human NKp30 protein has six isoforms. The canonical isoform (NKp30a) is 177 amino acids (aa) in length with a theoretical molecular weight of 21.6 kDa (1,2). The NKp30 protein consists of a signal peptide, an Ig domain, a stalk region, a transmembrane domain, and an intracellular region (1-3). NKp30/NCR3 signaling relies on interaction with the immunoreceptor tyrosine-based activation motif (ITAM)-containing adaptor molecules CD3zeta and FcRgamma (1,4,5). Other NRC family members include NKp44 and NKp46 (1,3-6). NK cell activation and tumor surveillance occurs through receptor-ligand binding. Tumor-derived ligands for NKp30 include HLA-B-associated transcript 3 (BAT3)/Bcl2-associated anthogene 6 (BAG6) and B7-H6 (4-6). NKp30 has also been suggested to play a role in NK-dendritic cell (DC) crosstalk by promoting DC maturation and cytotoxicity of immature DCs, mediated in part by BAT3 (1,4,5).

Given its role in NK cell-mediated killing and activation, NKp30 is a potential target for cancer immunotherapies (6). For instance, Bi-specific T cell engagers (BiTE) is one immunotherapy approach targeting the NKp30/B7-H6 interaction where B7-H6-specific BiTE elicits an attack on tumor cells through interferon gamma (IFNgamma) production and T cell cytotoxicity (1). Another potential approach is NKp30-derived chimeric antigen receptors (CARs) where CAR-T cells that express chimeric NKp30 receptors can efficiently attack B7-H6 tumor cells (1,6).

References

1. Barrow, A. D., Martin, C. J., & Colonna, M. (2019). The natural cytotoxicity receptors in health and disease. Frontiers in Immunology, 10, 909. https://doi.org/10.3389/fimmu.2019.00909

2. Uniprot (O14931)

3. Luczo, J. M., Ronzulli, S. L., & Tompkins, S. M. (2021). Influenza A virus hemagglutinin and other pathogen glycoprotein interactions with NK cell natural cytotoxicity receptors NKp46, NKp44, and NKp30. Viruses, 13(2), 156. https://doi.org/10.3390/v13020156

4. Pazina, T., Shemesh, A., Brusilovsky, M., Porgador, A., & Campbell, K. S. (2017). Regulation of the Functions of natural cytotoxicity receptors by interactions with diverse ligands and alterations in splice variant expression. Frontiers in Immunology, 8, 369.

5. Kaifu, T., Escaliere, B., Gastinel, L. N., Vivier, E., & Baratin, M. (2011). B7-H6/NKp30 interaction: a mechanism of alerting NK cells against tumors. Cellular and Molecular Life Sciences : CMLS, 68(21), 3531-3539. https://doi.org/10.1007/s00018-011-0802-7

6. Pinheiro, P. F., Justino, G. C., & Marques, M. M. (2020). NKp30 - A prospective target for new cancer immunotherapy strategies. British Journal of Pharmacology, 177(20), 4563-4580. https://doi.org/10.1111/bph.15222

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

Array NBL1-13521

• NKp30/NCR3 Antibodies
• NKp30/NCR3 Lysate
• NKp30/NCR3 Proteins and Enzymes