Aspirin-NHS Ester Mediated Acetylation of Lysine Residues in β-Conglycinin: Implications for Cancer Research in NF-κB

Post-translational modifications (PTMs) represent a dynamic and intricate layer of cellular regulation that significantly contributes to the diversification of protein functions (1). Among these modifications, lysine acetylation stands out as a crucial regulatory mechanism that modulates protein activities, subcellular localization, and interactions (2). This study focuses on expanding the role of lysine acetylation, a significant post-translational modification, within the nuclear factor-kappa B (NF-κB) transcription factor complex. NF-κB plays a crucial role in inflammation, immune responses, and cell survival, making it a pertinent target for cancer research (3). Dysregulation of NF-κB is implicated in various pathological conditions, emphasizing the need to explore lysine acetylation within this context. To perform the acetylation, we would attempt to use a common medication, Aspirin. Aspirin, or acetylsalicylic acid, is a widely studied non-steroidal anti-inflammatory drug (NSAID) that has been extensively used for its anti-inflammatory, analgesic, and antipyretic effects. In addition, the presence of an acetyl group on Aspirin results in a versatile molecule that can acetylate various proteins. (4). We synthesized Aspirin NHS ester (Aspirin-NHS) to capitalize on this versatility in acetylating biomolecules. This tailored approach allows for the selective targeting of lysine residues within proteins, offering a specific tool to examine the functional consequences of lysine acetylation within the NF-κB complex. For this study, a model protein system was used. We selected β-Conglycinin, a soy protein, that offers a pragmatic platform for investigating lysine acetylation within the NF-κB complex. β-Conglycinin is rich in lysine residues, similar to NF-κB, making it homologous to this line of inquiry (5). We can examine the theoretical considerations of lysine acetylation with practical experimental observations, providing a comprehensive understanding of the regulatory dynamics within the NF-κB complex. This research contributes not only to the expanding toolkit for precise protein acetylation modulation but also enhances our understanding of NF-κB's regulatory dynamics in the context of cancer, establishing a bridge between chemical synthesis and biological investigation.