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

The maintenance of machinery requires its operational understanding and a toolbox for repair. The methods for the precision engineering of native proteins meet a similar requirement in biosystems. Its success hinges on the principles regulating chemical reactions with a protein. Here, we report a te...

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Detalles Bibliográficos
Autores principales: Reddy, Neelesh C., Molla, Rajib, Joshi, Pralhad Namdev, T. K., Sajeev, Basu, Ipsita, Kawadkar, Jyotsna, Kalra, Neetu, Mishra, Ram Kumar, Chakrabarty, Suman, Shukla, Sanjeev, Rai, Vishal
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9561114/
https://www.ncbi.nlm.nih.gov/pubmed/36229616
http://dx.doi.org/10.1038/s41467-022-33772-1
Descripción
Sumario:The maintenance of machinery requires its operational understanding and a toolbox for repair. The methods for the precision engineering of native proteins meet a similar requirement in biosystems. Its success hinges on the principles regulating chemical reactions with a protein. Here, we report a technology that delivers high-level control over reactivity, chemoselectivity, site-selectivity, modularity, dual-probe installation, and protein-selectivity. It utilizes cysteine-based chemoselective Linchpin-Directed site-selective Modification of lysine residue in a protein (LDM(C-K)). The efficiency of the end-user-friendly protocol is evident in quantitative conversions within an hour. A chemically orthogonal C-S bond-formation and bond-dissociation are essential among multiple regulatory attributes. The method offers protein selectivity by targeting a single lysine residue of a single protein in a complex biomolecular mixture. The protocol renders analytically pure single-site probe-engineered protein bioconjugate. Also, it provides access to homogeneous antibody conjugates (AFC and ADC). The LDM(C-K)-ADC exhibits highly selective anti-proliferative activity towards breast cancer cells.