NEUROG3 Human

Neurogenin 3 (NGN3) is a basic helix-loop-helix (bHLH) transcription factor that plays a crucial role in the development of endocrine cells in the pancreas and intestine. It is also involved in the determination of neural precursor cells in the neuroectoderm. NGN3 is essential for the differentiation of endocrine progenitor cells into various hormone-producing cells, including insulin-producing β-cells. The recombinant form of Neurogenin 3 (Human Recombinant) is produced using advanced biotechnological methods to facilitate research and therapeutic applications.

The production of human recombinant Neurogenin 3 involves several steps to ensure high purity and biological activity. The gene encoding NGN3 is first codon-optimized to enhance protein expression in Escherichia coli (E. coli) strain BL21 (DE3). The codon-optimized NGN3 sequence is then fused in-frame to various fusion tags that facilitate cell penetration, nuclear translocation, and affinity purification .

The gene insert with the fusion tags is cloned into an expression vector, such as pET28a(+), for heterologous expression in E. coli cells. The expression conditions are optimized to produce a soluble form of the recombinant NGN3 fusion protein. The protein is then purified to homogeneity (purity >90%) under native conditions, retaining its secondary structure post-purification . This purified protein is tested for cytotoxicity and biological activity in human cells, ensuring its suitability for various biological applications .

Neurogenin 3 undergoes complex regulation, including post-translational modifications such as ubiquitylation. The ubiquitin proteasome system (UPS) is responsible for the rapid turnover of NGN3, regulating its stability and transcriptional activity. NGN3 can be ubiquitylated on lysines, the N-terminus, and non-canonical residues such as cysteines and serines/threonines . This rapid turnover is influenced by binding to its heterodimeric partner E protein and the presence of cyclin-dependent kinase (cdk) inhibitors .

The stability and activity of NGN3 are critical for its role in the specification and differentiation of endocrine cells. The regulation of NGN3 by the UPS and other post-translational mechanisms ensures precise control over its function during development and cellular processes .