Junming Ren, Alexander E Chu, Kevin M Jude, Lora K Picton, Aris J Kare, Leon Su, Alejandra Montano Romero, Po-Ssu Huang, K Christopher Garcia
Proceedings of the National Academy of Sciences of the United States of America 2022 Mar 22Affinity maturation of protein–protein interactions is an important approach in the development of therapeutic proteins such as cytokines. Typical experimental strategies involve targeting the cytokine-receptor interface with combinatorial libraries and then selecting for higher-affinity variants. Mutations to the binding scaffold are usually not considered main drivers for improved affinity. Here we demonstrate that computational design can provide affinity-enhanced variants of interleukin-2 (IL-2) “out of the box” without any requirement for interface engineering. Using a strategy of global IL-2 structural stabilization targeting metastable regions of the three-dimensional structure, rather than the receptor binding interfaces, we computationally designed thermostable IL-2 variants with up to 40-fold higher affinity for IL-2Rβ without any library-based optimization. These IL-2 analogs exhibited CD25-independent activities on T and natural killer (NK) cells both in vitro and in vivo, mimicking the properties of the IL-2 superkine “super-2” that was engineered through yeast surface display [A. M. Levin et al., Nature, 484, 529–533 (2012)]. Structure-guided stabilization of cytokines is a powerful approach to affinity maturation with applications to many cytokine and protein–protein interactions.
Junming Ren, Alexander E Chu, Kevin M Jude, Lora K Picton, Aris J Kare, Leon Su, Alejandra Montano Romero, Po-Ssu Huang, K Christopher Garcia. Interleukin-2 superkines by computational design. Proceedings of the National Academy of Sciences of the United States of America. 2022 Mar 22;119(12):e2117401119
PMID: 35294290
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