Rational engineering of lipid-binding probes via high-throughput protein-lipid interaction screening
NISHIMURA, T.; Tsuboyama, K.; Nakagaki, Y.; Yamamoto, E.; Mizushima, N.
Show abstract
Lipid-binding domains, originally isolated from natural proteins, are useful tools essential for analyzing membrane lipids in cells, and their applications are varied. Yet, there is no general strategy for engineering lipid-binding domains. Here, we present a robust method for monitoring protein-lipid interactions, named the Cell surface Liposome Binding (CLiB) assay. Using this technique, we isolated high-affinity lipid-binding domains and nanobodies that preferentially bind to phosphatidylinositol phosphates. Furthermore, by combining the CLiB assay with next-generation sequencing, allowing the analysis of more than 10,000 clones in parallel, we identified novel variants with enhanced binding to phosphatidylinositol phosphates and uncovered a common structural feature: a positively charged pocket necessary for binding, formed by the three loop regions. This study opens a new avenue for the rational design and generation of lipid-binding probes on demand.
Matching journals
The top 5 journals account for 50% of the predicted probability mass.
Similar papers in this journal
- Addressing lipid structural diversity in signalling: Photochemical probes for live-cell lipid biochemistry 95%
- ATG2A-mediated bridge-like lipid transport regulates lipid droplet accumulation 95%
- An In Vitro BRAF Activation Assay Elucidates Molecular Mechanisms Driving the Disassembly of the Autoinhibited BRAF State 95%
Similar papers in this journal
- Force redistribution in clathrin-mediated endocytosis revealed by coiled-coil force sensors 95%
- Ultrafast pore-loop dynamics in a AAA+ machine point to a Brownian-ratchet mechanism for protein translocation 94%
- Mechanistic insights into intramembrane proteolysis by E. coli site-2 protease homolog RseP 94%
Similar papers in this journal
- One-shot identification of SARS-CoV-2 S RBD escape mutants using yeast screening 93%
- The lysosomal membrane protein LAMP2B mediates microlipophagy to target obesity-related disorders 93%
- ERM-Dependent Assembly of T-Cell Receptor Signaling and Co-stimulatory Molecules on Microvilli Prior to Activation 93%
"Similar papers" are the closest papers from that journal in the model's embedding space. They show what the match is built on, but the ranking comes mostly from a classifier over the whole training set, not from these examples alone.