Welcome to the new version of CaltechAUTHORS. Login is currently restricted to library staff. If you notice any issues, please email coda@library.caltech.edu
Published April 6, 2024 | in press
Journal Article Open

Fast, accurate ranking of engineered proteins by target-binding propensity using structure modeling

  • 1. ROR icon California Institute of Technology

Abstract

Deep-learning-based methods for protein structure prediction have achieved unprecedented accuracy, yet their utility in the engineering of protein-based binders remains constrained due to a gap between the ability to predict the structures of candidate proteins and the ability toprioritize proteins by their potential to bind to a target. To bridge this gap, we introduce Automated Pairwise Peptide-Receptor Analysis for Screening Engineered proteins (APPRAISE), a method for predicting the target-binding propensity of engineered proteins. After generating structural models of engineered proteins competing for binding to a target using an established structure prediction tool such as AlphaFold-Multimer or ESMFold, APPRAISE performs a rapid (under 1 CPU second per model) scoring analysis that takes into account biophysical and geometrical constraints. As proof-of-concept cases, we demonstrate that APPRAISE can accurately classify receptor-dependent vs. receptor-independent adeno-associated viral vectors and diverse classes of engineered proteins such as miniproteins targeting the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike, nanobodies targeting a G-protein-coupled receptor, and peptides that specifically bind to transferrin receptor or programmed death-ligand 1 (PD-L1). APPRAISE is accessible through a web-based notebook interface using Google Colaboratory (https://tiny.cc/APPRAISE). With its accuracy, interpretability, and generalizability, APPRAISE promises to expand the utility of protein structure prediction and accelerate protein engineering for biomedical applications.

Copyright and License

© 2024 The Author(s) Under a Creative Commons license.

Acknowledgement

The authors thank Elisha Mackey, Zhe Qu, and Pat Anguiano for administrative assistance; Catherine Oikonomou for help with manuscript editing; and Sripriya R. Kumar, Seongmin Jang, Jimin Park, and Changfan Lin for helpful discussions. Schematics in this manuscript were created with BioRender.com. The study was funded by an NIH Director’s Pioneer Award DP1OD025535 (to V.G.). V.G. is the Director of Center for Molecular and Cellular Neuroscience of the Chen Institute at Caltech. All animal procedures in mice were approved by the California Institute of Technology Institutional Animal Care and Use Committee (IACUC), Caltech Office of Laboratory Animal Resources (OLAR), and were carried out in accordance with guidelines and regulations.

Contributions

X.D. and V.G. conceived the project. X.D. developed the APPRAISE method, applied APPRAISE to rank the engineered protein binders, and analyzed the results. X.C. identified PHP.D through directed evolution and characterized PHP.D in vivo. E.E.S. and T.F.S. designed and performed in vitro infectivity assays.

Conflict of Interest

V.G. is a scientific co-founder and BoD member of Capsida Biotherapeutics. The terms of the arrangements have been reviewed and approved by the California Institute of Technology in accordance with its conflict-of-interest policies.

Additional Information

During the preparation of this work, the authors used Stork Writing Assistant in order to improve grammar and writing clarity. After using this tool/service, the authors reviewed and edited the content as needed and take full responsibility for the content of the publication.

Data Availability

Document S1. Figures S1–S8 and Tables S1–S4

Files

1-s2.0-S1525001624002193-main.pdf
Files (9.7 MB)
Name Size Download all
md5:72822eff59dad244607fe80de2b4c379
5.2 MB Preview Download
md5:37550b379c8d2d96569e8a1f8393305c
4.5 MB Preview Download

Additional details

Created:
May 30, 2024
Modified:
June 4, 2024