MCPIP1/ZC3H12A 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.

Monocyte chemoattractant protein-induced protein 1 (MCPIP1), also referred to as ZC3H12A and Regnase-1, is an endonuclease that cleaves and destabilizes mRNA and functions as a negative regulator of inflammation (1,2). MCPIP1 is encoded by the ZC3H12A gene as a protein of 599 amino acids (aa) in length with a theoretical molecular weight of approximately 66 kDa (3-5). Common structural features of the MCPIP1 protein are the ubiquitin-associated (UBA) domain (43-89 aa), two protein-rich regions (PRRs) (100-126 aa, 458-536 aa), the PilT N-terminal (PIN) domain (133-270 aa), the zinc-finger motif (305-325 aa), the natively disordered region (326-457 aa), and the C-terminal conserved region (CCR) (545-598 aa) (1,3-5). MCPIP1 functions as an anti-inflammatory protein with RNase activity that is known to degrade many cytokine mRNA transcripts including interleukin-1beta (IL-1beta), IL-6, and IL-17 (4,5). In addition to its regulation of inflammation through mRNA degradation, MCPIP1 also controls inflammation through ubiquitin-specific peptidase 10 (USP10)-mediated protein deubiquitination of TRAF6 which results in suppression of c-Jun N-terminal kinase (JNK) and nuclear factor kappa B (NFkappaB) transcription factors (1,2,5). Besides regulating inflammation, MCPIP1 is involved in several non-immune cellular processes including apoptosis, adipogenesis, angiogenesis, and proliferation (2,5). MCPIP1 has also been implicated in a number of autoimmune diseases and other pathologies including chronic obstructive pulmonary disease (COPD), diabetes, rheumatoid arthritis, and cancer (3,5). Additionally, MCPIP1/ZC3H12A has been shown to have anti-viral activity against RNA viruses such as Japanese encephalitis virus and human immunodeficiency virus (HIV)-1 (1,5).

References

1. Uehata, T., & Takeuchi, O. (2017). Regnase-1 Is an Endoribonuclease Essential for the Maintenance of Immune Homeostasis. Journal of interferon & cytokine research: the official journal of the International Society for Interferon and Cytokine Research, 37(5), 220-229. https://doi.org/10.1089/jir.2017.0001

2. Miekus, K., Kotlinowski, J., Lichawska-Cieslar, A., Rys, J., & Jura, J. (2019). Activity of MCPIP1 RNase in tumor associated processes. Journal of experimental & clinical cancer research : CR, 38(1), 421. https://doi.org/10.1186/s13046-019-1430-6

3. Jura, J., Skalniak, L., & Koj, A. (2012). Monocyte chemotactic protein-1-induced protein-1 (MCPIP1) is a novel multifunctional modulator of inflammatory reactions. Biochimica et biophysica acta, 1823(10), 1905-1913. https://doi.org/10.1016/j.bbamcr.2012.06.029

4. Xu, J., Fu, S., Peng, W., & Rao, Z. (2012). MCP-1-induced protein-1, an immune regulator. Protein & cell, 3(12), 903-910. https://doi.org/10.1007/s13238-012-2075-9

5. Musson, R., Szukala, W., & Jura, J. (2020). MCPIP1 RNase and Its Multifaceted Role. International journal of molecular sciences, 21(19), 7183. https://doi.org/10.3390/ijms21197183

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 NBP2-06242

• MCPIP1/ZC3H12A Antibodies
• MCPIP1/ZC3H12A Lysate
• MCPIP1/ZC3H12A Proteins