C-Myc is a transcription factor encoded by the MYC gene located on human chromosome 8q24. It is a multifunctional, nuclear phosphoprotein that plays a crucial role in regulating cell growth, differentiation, metabolism, and apoptosis. The protein is part of the Myc family, which includes N-Myc and L-Myc, all of which share a basic-helix-loop-helix-leucine zipper (bHLHZip) domain . C-Myc is ubiquitously expressed in proliferating cells and is often dysregulated in various human cancers .
Recombinant C-Myc protein can be produced using various expression systems. One common method involves the use of an in vitro wheat germ expression system. This system is advantageous as it preserves the correct conformational folding necessary for the biological function of the protein . The recombinant protein is typically tagged with a GST (Glutathione S-transferase) tag at the N-terminal to facilitate purification and detection . The protein is then purified using affinity chromatography, and its purity is assessed by SDS-PAGE and Coomassie blue staining .
C-Myc functions as a transcription factor by forming a heterodimer with Max, another bHLHZip protein. This heterodimer binds to specific DNA sequences known as Enhancer-box (E-box) sequences (CANNTG) to regulate the transcription of target genes . The interaction between C-Myc and Max is crucial for the transcriptional activity of C-Myc. Additionally, C-Myc can interact with other proteins within the Myc/Max/Mxd network, influencing various cellular processes .
Mutations and overexpression of C-Myc are associated with uncontrolled cell proliferation and cancer . Therefore, understanding the biochemical interactions and regulatory mechanisms of C-Myc is essential for developing therapeutic strategies to inhibit its activity in cancer cells .