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A synthetic intrabody-based selective and generic inhibitor of GPCR endocytosis.

Nat Nanotechnol. 2017; 
Ghosh Eshan,Srivastava Ashish,Baidya Mithu,Kumari Punita,Dwivedi Hemlata,Nidhi Kumari,Ranjan Ravi,Dogra Shalini,Koide Akiko,Yadav Prem N,Sidhu Sachdev S,Koide Shohei,Shukla Ar
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Proteins, Expression, Isolation and Analysis Finally, the beads were washed 3–5 times with washing buffer as described above, and the eluted proteins were visualized by western blotting (βarr antibody (1:5,000 dilution (CST cat. no. 4674)); horseradish peroxidase (HRP)-coupled Protein L (1:2,000 dilution (GenScript cat. no. M00098)). Get A Quote

摘要

Beta-arrestins (βarrs) critically mediate desensitization, endocytosis and signalling of G protein-coupled receptors (GPCRs), and they scaffold a large number of interaction partners. However, allosteric modulation of their scaffolding abilities and direct targeting of their interaction interfaces to modulate GPCR functions selectively have not been fully explored yet. Here we identified a series of synthetic antibody fragments (Fabs) against different conformations of βarrs from phage display libraries. Several of these Fabs allosterically and selectively modulated the interaction of βarrs with clathrin and ERK MAP kinase. Interestingly, one of these Fabs selectively disrupted βarr-clathrin interac... More

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