Recombinant human TGF-β2 protein

Summary

• High purity human/bovine/porcine protein (UniProt: P61812)
• >98%, by SDS-PAGE quantitative densitometry
• Source: Expressed in E. coli
• 25.4 kDa dimer
• Animal origin-free (AOF) and carrier protein-free
• Manufactured in Cambridge, UK
• Lyophilized from acetonitrile/TFA
• Resuspend in 10 mM HCl, prepare single-use aliquots, add carrier protein if desired, and store frozen at -20°C or -80°C

Featured applications

• Induced pluripotent stem cell culture and maintenance
• Cell proliferation and differentiation
• Wound healing and tissue repair
• Induction of Epithelial-Mesenchymal Transition (EMT)
• Inflammatory response and immune modulation
• Stimulation of protein expression and secretion

Protein background

Transforming Growth Factor beta 2 (TGF-β2) belongs to theTGF-β superfamily, which encompasses TGF-β1, TGF-β2, and TGF-β3. These isoforms collectively regulate a wide array of cellular processes, including cell proliferation, differentiation, wound healing, apoptosis, and metabolism. The TGF-β superfamily signals through the same receptor, eliciting similar biological responses [1-4].

Overall, the TGF-β family plays pivotal roles in embryogenesis, tissue remodeling, and wound healing. TGF-β2’s significance in developmental processes is evident in mice with TGF-β2 deletions, displaying defects in cardiac, lung, craniofacial, limb, eye, ear, and urogenital systems [5]. It also exerts suppressive effects on IL-2-dependent T-cell growth, potentially enhancing tumour growth by suppressing immunosurveillance [5-6].

In cell culture applications, TGF-beta 2 is extensively employed due to its crucial regulatory roles. It modulates cell proliferation and differentiation by being added to the culture medium, regulating the growth and differentiation of various cell types, especially stem cells, providing a means to control and guide their fate into various cell types such as chondrocytes and epithelial-like cells [7-8].  For wound healing and tissue repair in cell culture models, TGF-beta 2 is utilized to drive cell migration, extracellular matrix production, and overall wound healing processes, creating a controlled environment to mimic conditions associated with tissue repair [3]. TGF-beta 2 is also a key inducer of epithelial-mesenchymal transition (EMT), allowing the study of cellular plasticity and transitions between different cell states [9-10].

TGF-β2’s involvement in immunomodulation, immune response suppression, and its association with tumour development make it valuable in investigating regulatory mechanisms of immune cells and understanding interactions within the tumour microenvironment [9,11]. Additionally, TGF-β2 stimulates the expression and secretion of specific proteins in cell culture, particularly relevant in studies focusing on extracellular matrix components. Researchers may combine TGF-β2 with other growth factors or cytokines to create a more physiologically relevant cellular environment in culture [11].

TGF-β2 is a 25.4 kDa protein composed of two identical 112 amino acid polypeptide chains linked by a single disulfide bond. It forms a latent complex with latency-associated peptide (LAP) and one of the latent TGF-β binding protein (LTBP) family members. This latent complex is stored at the cell surface and in the extracellular matrix. The release of biologically active TGF-β2 involves proteolytic processing of the complex and/or induction of conformational changes by proteins like thrombospondin-1, plasmin, matrix metalloproteases (MMP), or Integrins [12-13].

TGF-β2 signaling involves binding to a complex of the accessory receptor betaglycan (TGF-β RIII) and a type II ser/thr kinase receptor (TGF-β RII) and either TGF-β RI/ALK-5 or ALK-1. This complex activates Smad proteins that regulate transcription, with other Smad-independent pathways contributing to diverse cellular actions [4,14-15]

In summary, TGF-beta 2 plays a crucial role in regulating various cellular processes and is integral to the development of multiple organ systems, exhibiting diverse functions from controlling cell proliferation and differentiation to influencing immune responses and tumorigenesis.

Background references

1. Wakefield, L.et al.Regulation of transforming growth factor-β subtypes by members of the steroid hormone superfamily.Journal of Cell Science1990, 139–148 (1990).https://doi.org/10.1242/jcs.1990.supplement_13.13
2. Li, J.et al.TGF-β2 and TGF-β1 differentially regulate the odontogenic and osteogenic differentiation of mesenchymal stem cells.Archives of Oral Biology135, 105357 (2022).
3. Funaki, T.et al.Smad7 Suppresses the Inhibitory Effect of TGF-β2 on Corneal Endothelial Cell Proliferation and Accelerates Corneal Endothelial Wound Closure In Vitro:Cornea22, 153–159 (2003).
4. Shih, C.-C., Lee, C.-Y., Wong, F.-F. & Lin, C.-H. Protective Effects of One 2,4-Dihydro-3H-Pyrazol-3-one Derivative against Posterior Capsular Opacification by Regulation of TGF-β2/SMADs and Non-SMAD Signaling, Collagen I, and Fibronectin Proteins.CIMB44, 5048–5066 (2022).
5. Dünker, N. & Krieglstein, K. Targeted mutations of transforming growth factor‐β genes reveal important roles in mouse development and adult homeostasis.European Journal of Biochemistry267, 6982–6988 (2000).
6. Lee, H. K.et al.TGF-β2 antisense oligonucleotide enhances T-cell mediated anti-tumor activities by IL-2 via attenuation of fibrotic reaction in a humanized mouse model of pancreatic ductal adenocarcinoma.Biomedicine & Pharmacotherapy159, 114212 (2023).
7. Khan, N., Diaz-Hernandez, M. & Drissi, H. Differentiation of Human Induced Pluripotent Stem Cells (iPSCs)–derived Mesenchymal Progenitors into Chondrocytes.BIO-PROTOCOL13, (2023).
8. Yamashita, M., Aoki, H., Hashita, T., Iwao, T. & Matsunaga, T. Inhibition of transforming growth factor beta signaling pathway promotes differentiation of human induced pluripotent stem cell-derived brain microvascular endothelial-like cells.Fluids Barriers CNS17, 36 (2020).
9. Debnath, P.et al.Epithelial mesenchymal transition induced nuclear localization of the extracellular matrix protein Fibronectin.Biochimie219, 142–145 (2024).
10. Sim, H. J., Kim, M. R., Song, M. S. & Lee, S. Y. Kv3.4 regulates cell migration and invasion through TGF-β-induced epithelial–mesenchymal transition in A549 cells.Sci Rep14, 2309 (2024).
11. Salminen, A. AMPK signaling inhibits the differentiation of myofibroblasts: impact on age-related tissue fibrosis and degeneration.Biogerontology25, 83–106 (2024).
12. Oklü, R. & Hesketh, R. The latent transforming growth factor beta binding protein (LTBP) family.Biochem J352 Pt 3, 601–610 (2000).
13. Miyazono, K., Hellman, U., Wernstedt, C. & Heldin, C. H. Latent high molecular weight complex of transforming growth factor beta 1. Purification from human platelets and structural characterization.J Biol Chem263, 6407–6415 (1988).
14. De Caestecker, M. The transforming growth factor-β superfamily of receptors.Cytokine & Growth Factor Reviews15, 1–11 (2004).
15. Zúñiga, J. E.et al.Assembly of TbetaRI:TbetaRII:TGFbeta ternary complex in vitro with receptor extracellular domains is cooperative and isoform-dependent.J Mol Biol354, 1052–1068 (2005).