Defining short linear motif binding determinants by phage display‐based deep mutational scanning DOI Creative Commons
Caroline Benz,

Lars Maassen,

Leandro Simonetti

и другие.

Protein Science, Год журнала: 2025, Номер 34(6)

Опубликована: Май 24, 2025

Abstract Deep mutational scanning (DMS) has emerged as a powerful approach for evaluating the effects of mutations on binding or function. Here, we developed DMS by phage display protocol to define specificity determinants short linear motifs (SLiMs) peptide‐binding domains. We first designed benchmarking library evaluate performance well‐known ligands 11 different domains, including talin‐1 PTB domain, G3BP1 NTF2 and MDM2 SWIB domain. Comparison with set reference from eukaryotic motif (ELM) database confirmed that analysis correctly identifies known provides novel insights into determinants, defining non‐canonical putative extended conformation. A second was designed, aiming provide information 19 SLiM‐based interactions between human SARS‐CoV‐2 proteins. The affinity determining residues viral peptides host proteins refined consensus in five domains proteins, non‐structural protein (NSP) 9. further pinpointed increased NSP3 NSP9. An affinity‐improved cell‐permeable NSP9‐binding peptide found exert stronger antiviral than wild‐type peptide. Our study demonstrates can efficiently be multiplexed applied refine shows how results guide peptide‐engineering efforts.

Язык: Английский

Defining short linear motif binding determinants by phage display‐based deep mutational scanning DOI Creative Commons
Caroline Benz,

Lars Maassen,

Leandro Simonetti

и другие.

Protein Science, Год журнала: 2025, Номер 34(6)

Опубликована: Май 24, 2025

Abstract Deep mutational scanning (DMS) has emerged as a powerful approach for evaluating the effects of mutations on binding or function. Here, we developed DMS by phage display protocol to define specificity determinants short linear motifs (SLiMs) peptide‐binding domains. We first designed benchmarking library evaluate performance well‐known ligands 11 different domains, including talin‐1 PTB domain, G3BP1 NTF2 and MDM2 SWIB domain. Comparison with set reference from eukaryotic motif (ELM) database confirmed that analysis correctly identifies known provides novel insights into determinants, defining non‐canonical putative extended conformation. A second was designed, aiming provide information 19 SLiM‐based interactions between human SARS‐CoV‐2 proteins. The affinity determining residues viral peptides host proteins refined consensus in five domains proteins, non‐structural protein (NSP) 9. further pinpointed increased NSP3 NSP9. An affinity‐improved cell‐permeable NSP9‐binding peptide found exert stronger antiviral than wild‐type peptide. Our study demonstrates can efficiently be multiplexed applied refine shows how results guide peptide‐engineering efforts.

Язык: Английский

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