Object's details: Next-generation sequencing of a combinatorial peptide phage library screened against ubiquitin identifies peptide aptamers that can inhibit the in vitro ubiquitin transfer cascade

Provider:Repozytorium Instytutu Biochemii i Biofizyki Polskiej Akademii Nauk

Description

Title:
Next-generation sequencing of a combinatorial peptide phage library screened against ubiquitin identifies peptide aptamers that can inhibit the in vitro ubiquitin transfer cascade
Creator:
Lisowska, Małgorzata;
Lickiss, Fiona;
Gil-Mir, Maria;
Huart, Anne-Sophie;
Trybala, Zuzanna;
Way, Luke;
Hernychova, Lenka;
Krejci, Adam;
Muller, Petr;
Krejcir, Radovan;
Zhukov, Igor;
Jurczak, Przemyslaw;
Rodziewicz-Motowidło, Sylwia;
Ball, Kathryn;
Vojtesek, Borivoj;
Hupp, Ted;
Kalathiya, Umesh
Description:
Defining dynamic protein–protein interactions in the ubiquitin conjugation reaction is a challenging research area. Generating peptide aptamers that target components such as ubiquitin itself, E1, E2, or E3 could provide tools to dissect novel features of the enzymatic cascade. Next-generation deep sequencing platforms were used to identify peptide sequences isolated from phage-peptide libraries screened against Ubiquitin and its ortholog NEDD8. In over three rounds of selection under differing wash criteria, over 13,000 peptides were acquired targeting ubiquitin, while over 10,000 peptides were selected against NEDD8. The overlap in peptides against these two proteins was less than 5% suggesting a high degree in specificity of Ubiquitin or NEDD8 toward linear peptide motifs. Two of these ubiquitin-binding peptides were identified that inhibit both E3 ubiquitin ligases MDM2 and CHIP. NMR analysis highlighted distinct modes of binding of the two different peptide aptamers. These data highlight the utility of using next-generation sequencing of combinatorial phage-peptide libraries to isolate peptide aptamers toward a protein target that can be used as a chemical tool in a complex multi-enzyme reaction.
Object availability:
free access
Date:
2022.12.02
Type:
Article;
PeerReviewed
Language:
English;
eng
Format:
application/pdf
Publisher:
Frontiers
Relation: http://eprints.ibb.waw.pl/2218 ;http://doi.org/10.3389/fmicb.2022.875556 ;doi:10.3389/fmicb.2022.875556
Subject:
Q Science (General);
QD Chemistry;
QR Microbiology
Identifier: http://eprints.ibb.waw.pl/2218/1/frontiers.in.microbiology_2022_13_875556.pdf ;
Lisowska, Małgorzata and Lickiss, Fiona and Gil-Mir, Maria and Huart, Anne-Sophie and Trybala, Zuzanna and Way, Luke and Hernychova, Lenka and Krejci, Adam and Muller, Petr and Krejcir, Radovan and Zhukov, Igor and Jurczak, Przemyslaw and Rodziewicz-Motowidło, Sylwia and Ball, Kathryn and Vojtesek, Borivoj and Hupp, Ted and Kalathiya, Umesh (2022) Next-generation sequencing of a combinatorial peptide phage library screened against ubiquitin identifies peptide aptamers that can inhibit the in vitro ubiquitin transfer cascade. Frontiers in Microbiology, 13 . .875556. ISSN 1664-302X
Data provider:
Repozytorium Instytutu Biochemii i Biofizyki Polskiej Akademii Nauk

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