Engineering an autonomous VH domain to modulate intracellular pathways and to interrogate the eIF4F complex.
- Resource Type
- Academic Journal
- Authors
- Frosi Y; p53 Laboratory (ASTAR), 8A Biomedical Grove, #06-04/05, Neuros/Immunos, 138648, Singapore.; Disease Intervention Technology Laboratory (DITL), Institute of Molecular and Cell Biology, ASTAR, Singapore, 138673, Singapore.; Lin YC; p53 Laboratory (ASTAR), 8A Biomedical Grove, #06-04/05, Neuros/Immunos, 138648, Singapore.; Insilico Medicine Taiwan Ltd., Taipei City, 110208, Taiwan.; Department of Pharmacy, National Yang Ming Chiao Tung University, Taipei City, 112304, Taiwan.; Shimin J; p53 Laboratory (ASTAR), 8A Biomedical Grove, #06-04/05, Neuros/Immunos, 138648, Singapore.; Disease Intervention Technology Laboratory (DITL), Institute of Molecular and Cell Biology, ASTAR, Singapore, 138673, Singapore.; Ramlan SR; p53 Laboratory (ASTAR), 8A Biomedical Grove, #06-04/05, Neuros/Immunos, 138648, Singapore.; Disease Intervention Technology Laboratory (DITL), Institute of Molecular and Cell Biology, ASTAR, Singapore, 138673, Singapore.; Hew K; DotBio Pte. Ltd., 1 Research Link, Singapore, 117604, Singapore.; School of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive, 637551, Singapore, Singapore.; Engman AH; DotBio Pte. Ltd., 1 Research Link, Singapore, 117604, Singapore.; School of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive, 637551, Singapore, Singapore.; Pillai A; DotBio Pte. Ltd., 1 Research Link, Singapore, 117604, Singapore.; School of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive, 637551, Singapore, Singapore.; Yeung K; DotBio Pte. Ltd., 1 Research Link, Singapore, 117604, Singapore.; School of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive, 637551, Singapore, Singapore.; Cheng YX; DotBio Pte. Ltd., 1 Research Link, Singapore, 117604, Singapore.; School of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive, 637551, Singapore, Singapore.; Cornvik T; School of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive, 637551, Singapore, Singapore.; Nordlund P; DotBio Pte. Ltd., 1 Research Link, Singapore, 117604, Singapore.; School of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive, 637551, Singapore, Singapore.; Department of Oncology and Pathology, Karolinska Institutet, Stockholm, 17177, Sweden.; Goh M; p53 Laboratory (ASTAR), 8A Biomedical Grove, #06-04/05, Neuros/Immunos, 138648, Singapore.; Lama D; Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, Biomedicum Quarter 7B-C Solnavägen 9, 17165, Solna, Sweden.; Gates ZP; Disease Intervention Technology Laboratory (DITL), Institute of Molecular and Cell Biology, ASTAR, Singapore, 138673, Singapore.; Institute of Sustainability for Chemicals, Energy and Environment (ISCE2), ASTAR, 8 A Biomedical Grove, #07-01 Neuros Building, 138665, Singapore, Singapore.; Verma CS; School of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive, 637551, Singapore, Singapore.; Bioinformatics Institute (ASTAR), 30 Biopolis Street, #07-01 Matrix, 138671, Singapore, Singapore.; Department of Biological Sciences, National University of Singapore, 14 Science Drive 4, 117543, Singapore, Singapore.; Thean D; p53 Laboratory (ASTAR), 8A Biomedical Grove, #06-04/05, Neuros/Immunos, 138648, Singapore.; Lane DP; p53 Laboratory (ASTAR), 8A Biomedical Grove, #06-04/05, Neuros/Immunos, 138648, Singapore.; Asial I; DotBio Pte. Ltd., 1 Research Link, Singapore, 117604, Singapore. ignacio.asial@dotbiopharma.com.; School of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive, 637551, Singapore, Singapore. ignacio.asial@dotbiopharma.com.; Brown CJ; p53 Laboratory (ASTAR), 8A Biomedical Grove, #06-04/05, Neuros/Immunos, 138648, Singapore. cjbrown@imcb.a-star.edu.sg.; Disease Intervention Technology Laboratory (DITL), Institute of Molecular and Cell Biology, ASTAR, Singapore, 138673, Singapore. cjbrown@imcb.a-star.edu.sg.
- Source
- Publisher: Nature Pub. Group Country of Publication: England NLM ID: 101528555 Publication Model: Electronic Cited Medium: Internet ISSN: 2041-1723 (Electronic) Linking ISSN: 20411723 NLM ISO Abbreviation: Nat Commun Subsets: MEDLINE
- Subject
- Language
- English
An attractive approach to target intracellular macromolecular interfaces and to model putative drug interactions is to design small high-affinity proteins. Variable domains of the immunoglobulin heavy chain (VH domains) are ideal miniproteins, but their development has been restricted by poor intracellular stability and expression. Here we show that an autonomous and disufhide-free VH domain is suitable for intracellular studies and use it to construct a high-diversity phage display library. Using this library and affinity maturation techniques we identify VH domains with picomolar affinity against eIF4E, a protein commonly hyper-activated in cancer. We demonstrate that these molecules interact with eIF4E at the eIF4G binding site via a distinct structural pose. Intracellular overexpression of these miniproteins reduce cellular proliferation and expression of malignancy-related proteins in cancer cell lines. The linkage of high-diversity in vitro libraries with an intracellularly expressible miniprotein scaffold will facilitate the discovery of VH domains suitable for intracellular applications.
(© 2022. The Author(s).)