Development of a multipurpose scaffold for the display of peptide loops

Maxim Rossman; Sandra Joy Muriatu Greive; Tommasso Moschetti; Michael Dinan; Marko Hyvonen

doi:10.1093/protein/gzx017

Development of a multipurpose scaffold for the display of peptide loops

Maxim Rossman, Sandra Joy Muriatu Greive, Tommasso Moschetti, Michael Dinan, Marko Hyvonen

Chemistry

Research output: Contribution to journal › Article › peer-review

Abstract

Protein–protein interactions (PPIs) determine a wide range of biological processes and analysis of these dynamic networks is increasingly becoming a mandatory tool for studying protein function. Using the globular ATPase domain of recombinase RadA as a scaffold, we have developed a peptide display system (RAD display), which allows for the presentation of target peptides, protein domains or full-length proteins and their rapid recombinant production in bacteria. The design of the RAD display system includes differently tagged versions of the scaffold, which allows for flexibility in the protein purification method, and chemical coupling for small molecule labeling or surface immobilization. When combined with the significant thermal stability of the RadA protein, these features create a versatile multipurpose scaffold system. Using various orthogonal biophysical techniques, we show that peptides displayed on the scaffold bind to their natural targets in a fashion similar to linear parent peptides. We use the examples of CK2β/CK2α kinase and TPX2/Aurora A kinase protein complexes to demonstrate that the peptide displayed by the RAD scaffold can be used in PPI studies with the same binding efficacy but at lower costs compared with their linear synthetic counterparts.

Original language	English
Pages (from-to)	419-430
Number of pages	12
Journal	Protein Engineering, Design and Selection
Volume	30
Issue number	6
DOIs	https://doi.org/10.1093/protein/gzx017
Publication status	Published - 24 Apr 2017

Bibliographical note

Access to Document

10.1093/protein/gzx017

gzx017
© The Author 2017. Published by Oxford University Press.
Final published version, 668 KBLicence: CC BY

Cite this

@article{7389917170af4e96921ecb57f7520faf,

title = "Development of a multipurpose scaffold for the display of peptide loops",

abstract = "Protein–protein interactions (PPIs) determine a wide range of biological processes and analysis of these dynamic networks is increasingly becoming a mandatory tool for studying protein function. Using the globular ATPase domain of recombinase RadA as a scaffold, we have developed a peptide display system (RAD display), which allows for the presentation of target peptides, protein domains or full-length proteins and their rapid recombinant production in bacteria. The design of the RAD display system includes differently tagged versions of the scaffold, which allows for flexibility in the protein purification method, and chemical coupling for small molecule labeling or surface immobilization. When combined with the significant thermal stability of the RadA protein, these features create a versatile multipurpose scaffold system. Using various orthogonal biophysical techniques, we show that peptides displayed on the scaffold bind to their natural targets in a fashion similar to linear parent peptides. We use the examples of CK2β/CK2α kinase and TPX2/Aurora A kinase protein complexes to demonstrate that the peptide displayed by the RAD scaffold can be used in PPI studies with the same binding efficacy but at lower costs compared with their linear synthetic counterparts.",

author = "Maxim Rossman and Greive, {Sandra Joy Muriatu} and Tommasso Moschetti and Michael Dinan and Marko Hyvonen",

note = "{\textcopyright} The Author 2017. Published by Oxford University Press.",

year = "2017",

month = apr,

day = "24",

doi = "10.1093/protein/gzx017",

language = "English",

volume = "30",

pages = "419--430",

journal = "Protein Engineering, Design and Selection",

issn = "1741-0126",

publisher = "Oxford University Press",

number = "6",

}

TY - JOUR

T1 - Development of a multipurpose scaffold for the display of peptide loops

AU - Rossman, Maxim

AU - Greive, Sandra Joy Muriatu

AU - Moschetti, Tommasso

AU - Dinan, Michael

AU - Hyvonen, Marko

PY - 2017/4/24

Y1 - 2017/4/24

N2 - Protein–protein interactions (PPIs) determine a wide range of biological processes and analysis of these dynamic networks is increasingly becoming a mandatory tool for studying protein function. Using the globular ATPase domain of recombinase RadA as a scaffold, we have developed a peptide display system (RAD display), which allows for the presentation of target peptides, protein domains or full-length proteins and their rapid recombinant production in bacteria. The design of the RAD display system includes differently tagged versions of the scaffold, which allows for flexibility in the protein purification method, and chemical coupling for small molecule labeling or surface immobilization. When combined with the significant thermal stability of the RadA protein, these features create a versatile multipurpose scaffold system. Using various orthogonal biophysical techniques, we show that peptides displayed on the scaffold bind to their natural targets in a fashion similar to linear parent peptides. We use the examples of CK2β/CK2α kinase and TPX2/Aurora A kinase protein complexes to demonstrate that the peptide displayed by the RAD scaffold can be used in PPI studies with the same binding efficacy but at lower costs compared with their linear synthetic counterparts.

AB - Protein–protein interactions (PPIs) determine a wide range of biological processes and analysis of these dynamic networks is increasingly becoming a mandatory tool for studying protein function. Using the globular ATPase domain of recombinase RadA as a scaffold, we have developed a peptide display system (RAD display), which allows for the presentation of target peptides, protein domains or full-length proteins and their rapid recombinant production in bacteria. The design of the RAD display system includes differently tagged versions of the scaffold, which allows for flexibility in the protein purification method, and chemical coupling for small molecule labeling or surface immobilization. When combined with the significant thermal stability of the RadA protein, these features create a versatile multipurpose scaffold system. Using various orthogonal biophysical techniques, we show that peptides displayed on the scaffold bind to their natural targets in a fashion similar to linear parent peptides. We use the examples of CK2β/CK2α kinase and TPX2/Aurora A kinase protein complexes to demonstrate that the peptide displayed by the RAD scaffold can be used in PPI studies with the same binding efficacy but at lower costs compared with their linear synthetic counterparts.

U2 - 10.1093/protein/gzx017

DO - 10.1093/protein/gzx017

M3 - Article

VL - 30

SP - 419

EP - 430

JO - Protein Engineering, Design and Selection

JF - Protein Engineering, Design and Selection

SN - 1741-0126

IS - 6

ER -