biostudies-literatureRen JG. Harold and Leila Y. Mathers Charitable FoundationHHS | National Institutes of HealthU.S. Department of EnergyHoward Hughes Medical InstituteNIAID NIH HHSLudwig Institute for Cancer ResearchNIGMS NIH HHSe2117401119https://www.ebi.ac.uk/biostudies/studies/S-EPMC8944926biostudies-literatureUnknown119(12)Affinity 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 Proceedings of the National Academy of Sciences of the United States of AmericaInterleukin-2 superkines by computational design.PMC8944926n/aR37 AI051321P30 GM133894R01 AI051321SciDACNIH-RO1-AI51321MF-1802-00128Picton LKGarcia KCJude KMRen JChu AEHuang PSMontano Romero ASu LKare AJfalseInterleukin-2 superkines by computational design.Affinity 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 2022-01-01T00:00:00Z2022 Mar2026-05-30T18:48:12.222Z2024-12-04T03:40:27.49ZS-EPMC89449263529429010.1073/pnas.2117401119