Recombinant mouse small proline-rich protein 2A1 (Sprr2a1) production in E. coli involves co-inserting the gene of interest (1-83aa of mouse Sprr2a1) into an expression vector with an N-terminal 6xHis-tag gene and transforming it into E. coli cells. The cells are cultured under conditions that induce protein expression. After sufficient growth, the cells are lysed to release the recombinant Sprr2a1 protein. The obtained recombinant Sprr2a1 protein is purified through the affinity chromatography technique. The purity of the Sprr2a1 protein is assessed using SDS-PAGE, exceeding 90%.
SPRR2a1 is a member of the small proline-rich protein (SPRR) family, which is essential for forming the cornified cell envelope (CE) in various tissues, such as the skin and uterus. The CE serves as a protective barrier against environmental stress and dehydration [1]. SPRR2a1 is implicated in allergic inflammation and is upregulated in response to IL-6/STAT3 signaling [2]. Studies have demonstrated that SPRR proteins are closely associated with genes encoding other structural proteins like keratin and are vital for maintaining barrier function in the skin [3][4].
Moreover, research has shown that estrogen regulates SPRR2a1 in the mouse uterus [5]. The expression of SPRR proteins, including SPRR2a1, is influenced by hormonal fluctuations during the estrous cycle and pregnancy [5]. This underscores the significance of SPRR proteins in the female reproductive system.
References:
[1] H. Lee, H. Kim, Y. Lee, S. Lee, W. Kim, S. Hanet al., Immunolocalisation and oestrogen regulation of small proline-rich protein 2a protein in the mouse uterus, Reproduction Fertility and Development, vol. 26, no. 5, p. 682, 2014. https://doi.org/10.1071/rd12408
[2] I. Nozaki, J. Lunz, S. Specht, D. Stolz, K. Taguchi, В. Субботинet al., Small proline-rich proteins 2 are noncoordinately upregulated by il-6/stat3 signaling after bile duct ligation, Laboratory Investigation, vol. 85, no. 1, p. 109-123, 2005. https://doi.org/10.1038/labinvest.3700213
[3] J. Segre, Epidermal barrier formation and recovery in skin disorders, Journal of Clinical Investigation, vol. 116, no. 5, p. 1150-1158, 2006. https://doi.org/10.1172/jci28521
[4] N. Zimmermann, M. Doepker, D. Witte, K. Stringer, P. Fulkerson, S. Popeet al., Expression and regulation of small proline-rich protein 2 in allergic inflammation, American Journal of Respiratory Cell and Molecular Biology, vol. 32, no. 5, p. 428-435, 2005. https://doi.org/10.1165/rcmb.2004-0269oc
[5] Y. Tan, X. Sun, F. Li, S. Tang, Y. Piao, & Y. Wang, Gene expression pattern and hormonal regulation of small proline-rich protein 2 family members in the female mouse reproductive system during the estrous cycle and pregnancy, Reproduction Nutrition Development, vol. 46, no. 6, p. 641-655, 2006. https://doi.org/10.1051/rnd:2006037
