%0 Journal Article %1 schulte2021highthroughput %A Schulte, Clemens %A Khayenko, Vladimir %A Nordblom, Noah Frieder %A Tippel, Franziska %A Peck, Violetta %A Gupta, Amit Jean %A Maric, Hans Michael %D 2021 %J iScience %K maric %N 1 %P 101898-- %R https://doi.org/10.1016/j.isci.2020.101898 %T High-throughput determination of protein affinities using unmodified peptide libraries in nanomolar scale %U https://www.sciencedirect.com/science/article/pii/S2589004220310956 %V 24 %X Protein-protein interactions (PPIs) are of fundamental importance for our understanding of physiology and pathology. PPIs involving short, linear motifs play a major role in immunological recognition, signaling, and regulation and provide attractive starting points for pharmaceutical intervention. Yet, state-of-the-art protein-peptide affinity determination approaches exhibit limited throughput and sensitivity, often resulting from ligand immobilization, labeling, or synthesis. Here, we introduce a high-throughput method for in-solution analysis of protein-peptide interactions using a phenomenon called temperature related intensity change (TRIC). We use TRIC for the identification and fine-mapping of low- and high-affinity protein interaction sites and the definition of sequence binding requirements. Validation is achieved by microarray-based studies using wild-type and mutated recombinant protein and the native protein within tissue lysates. On-chip neutralization and strong correlation with structural data establish TRIC as a quasi-label-free method to determine binding affinities of unmodified peptide libraries with large dynamic range.

@article{schulte2021highthroughput, abstract = {Protein-protein interactions (PPIs) are of fundamental importance for our understanding of physiology and pathology. PPIs involving short, linear motifs play a major role in immunological recognition, signaling, and regulation and provide attractive starting points for pharmaceutical intervention. Yet, state-of-the-art protein-peptide affinity determination approaches exhibit limited throughput and sensitivity, often resulting from ligand immobilization, labeling, or synthesis. Here, we introduce a high-throughput method for in-solution analysis of protein-peptide interactions using a phenomenon called temperature related intensity change (TRIC). We use TRIC for the identification and fine-mapping of low- and high-affinity protein interaction sites and the definition of sequence binding requirements. Validation is achieved by microarray-based studies using wild-type and mutated recombinant protein and the native protein within tissue lysates. On-chip neutralization and strong correlation with structural data establish TRIC as a quasi-label-free method to determine binding affinities of unmodified peptide libraries with large dynamic range.}, added-at = {2021-04-07T09:23:02.000+0200}, author = {Schulte, Clemens and Khayenko, Vladimir and Nordblom, Noah Frieder and Tippel, Franziska and Peck, Violetta and Gupta, Amit Jean and Maric, Hans Michael}, biburl = {https://www.bibsonomy.org/bibtex/2d011c9461aa23af77bb7a0754730e7d5/reichert}, doi = {https://doi.org/10.1016/j.isci.2020.101898}, interhash = {9b329b851697a7546b7ee7b386f5b0ae}, intrahash = {d011c9461aa23af77bb7a0754730e7d5}, issn = {25890042}, journal = {iScience}, keywords = {maric}, number = 1, pages = {101898--}, timestamp = {2021-04-07T09:23:02.000+0200}, title = {High-throughput determination of protein affinities using unmodified peptide libraries in nanomolar scale}, url = {https://www.sciencedirect.com/science/article/pii/S2589004220310956}, volume = 24, year = 2021 }