Traceless cysteine-linchpin enables precision engineering of lysine in native proteins

• DOI: 10.1038/s41467-022-33772-1
• Publication Date: 2022-10-13T10:02:53Z
• Authors: Neelesh C. Reddy; Rajib Molla; Pralhad Namdev Joshi; Sajeev T. K.; Ipsita Basu; Jyotsna Kawadkar; Neetu Kalra; Ram Kumar Mishra; Suman Chakrabarty; Sanjeev Shukla; Vishal Rai
• Keywords: Protein Engineering

Abstract The maintenance of machinery requires its operational understanding and a toolbox for repair. methods the precision engineering native proteins meet similar requirement in biosystems. Its success hinges on principles regulating chemical reactions with protein. Here, we report technology that delivers high-level control over reactivity, chemoselectivity, site-selectivity, modularity, dual-probe installation, protein-selectivity. It utilizes cysteine-based chemoselective Linchpin-Directed site-selective Modification lysine residue protein (LDM C-K ). efficiency end-user-friendly protocol is evident quantitative conversions within an hour. A chemically orthogonal C-S bond-formation bond-dissociation are essential among multiple regulatory attributes. method offers selectivity by targeting single complex biomolecular mixture. renders analytically pure single-site probe-engineered bioconjugate. Also, it provides access to homogeneous antibody conjugates (AFC ADC). LDM -ADC exhibits highly selective anti-proliferative activity towards breast cancer cells.