To produce recombinant human protein kinase C eta type (PRKCH) in E. coli, the full-length gene fragment of the PRKCH protein (1-683aa) is co-inserted into an expression vector with a C-terminal 6xHis-tag gene and transformed into E. coli cells. The bacteria are cultured under conditions that promote protein expression. After reaching sufficient growth, the cells are lysed to release the recombinant PRKCH protein. Purification is done using affinity chromatography. The purity of the PRKCH protein is confirmed using SDS-PAGE, exceeding 90%.
PRKCH belongs to the Ca2+-independent novel PKC subfamily and is located on human chromosome 14 and mouse chromosome 12 [1]. PRKCH is unique within the PKC family due to its distinct regulation compared to other PKC isozymes [2]. It is highly abundant in T cells and plays a crucial role in T cell activation and homeostatic proliferation [3]. Additionally, PRKCH is necessary for T-cell activation and regulatory T-cell function [4]. Furthermore, PRKCH has been implicated in keratinocyte growth and differentiation control [5].
PRKCH is known to be involved in various cellular functions and processes. For instance, it has been shown to regulate hematopoietic stem cell function and predict poor prognosis in acute myeloid leukemia [4]. Moreover, PRKCH is required for keratinocyte growth arrest and differentiation [5]. In lung tumors, PRKCH has been identified with a unique tissue distribution in the skin and lungs [6].
References:
[1] M. Kashiwagi, M. Ohba, K. Chida, & T. Kuroki, Protein kinase c (pkc ): its involvement in keratinocyte differentiation, The Journal of Biochemistry, vol. 132, no. 6, p. 853-857, 2002. https://doi.org/10.1093/oxfordjournals.jbchem.a003297
[2] A. Basu, The enigmatic protein kinase c-eta, Cancers, vol. 11, no. 2, p. 214, 2019. https://doi.org/10.3390/cancers11020214
[3] G. Fu, J. Hu, N. Niederberger-Magnenat, V. Rybakin, J. Casas, P. Yachiet al., Protein kinase c η is required for t cell activation and homeostatic proliferation, Science Signaling, vol. 4, no. 202, 2011. https://doi.org/10.1126/scisignal.2002058
[4] S. Porter and J. Magee, Prkch regulates hematopoietic stem cell function and predicts poor prognosis in acute myeloid leukemia, Experimental Hematology, vol. 53, p. 43-47, 2017. https://doi.org/10.1016/j.exphem.2017.05.006
[5] S. Cabodi, E. Calautti, C. Talora, T. Kuroki, P. Stein, & G. Dotto, A pkc-η/fyn-dependent pathway leading to keratinocyte growth arrest and differentiation, Molecular Cell, vol. 6, no. 5, p. 1121-1129, 2000. https://doi.org/10.1016/s1097-2765(00)00110-6
[6] F. Canzian, M. Gariboldi, G. Manenti, L. Gregorio, S. Osada, S. Ohnoet al., Expression in lung tumors and genetic mapping of the novel murine protein kinase cη, Molecular Carcinogenesis, vol. 9, no. 3, p. 111-113, 1994. https://doi.org/10.1002/mc.2940090302
