The production of recombinant human Thymosin beta-4 (TMSB4X) begins with isolating the gene encoding the TMSB4X protein (2-44aa). This gene is cloned into an expression vector and introduced into E. coli cells. The E. coli cells are cultured in bioreactors to express the protein. After the cells have grown sufficiently, they are harvested and lysed to release the TMSB4X protein. The TMSB4X protein is purified using affinity chromatography. Finally, rigorous quality control tests are conducted to ensure the protein's purity and functionality, making it suitable for research use. Its activity has been confirmed by its ability to produce a protective effect against hydrogen peroxide in primary lung fibroblasts in a concentration range of 0.5 - 10 μg/ml. Its endotoxin content is less than 1.0 EU/μg as determined by the LAL method. Its purity is greater than 97% as determined by SDS-PAGE.
TMSB4X is an actin-sequestering protein involved in various cellular processes, including regulation of actin polymerization, cytoskeleton organization, cell proliferation, migration, differentiation, wound healing, apoptosis, and cell survival [1][2][3]. TMSB4X is crucial for adherens junction stability, epidermal planar cell polarity, and the morphogenesis and differentiation of coronary vessels during embryonic development [4]. TMSB4X has been associated with early preneoplastic cell transformation in pancreatic cancer and plays a role in EMT induction and metastasis through the TGF-ß/Tß4/MRTF signaling pathway [5][6].
References:
[1] L. Hayden, B. Hobbs, R. Busch, M. Cho, M. Liu, C. Lopes-Ramoset al., X chromosome associations with chronic obstructive pulmonary disease and related phenotypes: an x chromosome-wide association study, Respiratory Research, vol. 24, no. 1, 2023. https://doi.org/10.1186/s12931-023-02337-1
[2] D. Rengaraj, Y. Hwang, X. Liang, W. Deng, Z. Yang, & J. Han, Comparative expression and regulation of tmsb4x in male reproductive tissues of rats and chickens, Journal of Experimental Zoology Part a Ecological Genetics and Physiology, vol. 319, no. 10, p. 584-595, 2013. https://doi.org/10.1002/jez.1820
[3] M. Gezen, U. Yilmaz, O. Fazlıoğulları, N. Yilmaz, C. Horozoglu, A. Ergenet al., Thymosin beta-4 a/t polymorphism and acute coronary syndrome risk, Archives of Clinical and Experimental Medicine, vol. 4, no. 1, p. 17-20, 2019. https://doi.org/10.25000/acem.475094
[4] K. Padmanabhan, H. Grobe, J. Cohen, A. Soffer, A. Mahly, O. Adiret al., Thymosin β4 is essential for adherens junction stability and epidermal planar cell polarity, Development, vol. 147, no. 23, 2020. https://doi.org/10.1242/dev.193425
[5] B. Grüner, H. Hahne, P. Mazur, M. Trajkovic‐Arsic, S. Maier, I. Espósitoet al., Maldi imaging mass spectrometry for in situ proteomic analysis of preneoplastic lesions in pancreatic cancer, Plos One, vol. 7, no. 6, p. e39424, 2012. https://doi.org/10.1371/journal.pone.0039424
[6] R. Moradpoor, A. Gharebaghian, F. Shahi, A. Mousavi, S. Salari, M. Akbariet al., Identification and validation of stage-associated pbmc biomarkers in breast cancer using ms-based proteomics, Frontiers in Oncology, vol. 10, 2020. https://doi.org/10.3389/fonc.2020.01101
