A Multivalent Structure‐Specific RNA Binder with Extremely Stable Target Binding but Reduced Interaction with Nonspecific RNAs

By greatly enhancing binding affinities against target biomolecules, multivalent interactions provide an attractive strategy for biosensing. However, there is also a major concern for increased binding to nonspecific targets by multivalent binding. A range of charge‐engineered probes of a structure‐...

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Bibliographic Details
Published in:Angewandte Chemie International Edition 2017-12, Vol.56 (50), p.15998-16002
Main Authors: Lee, Jeong Min, Hwang, Ahreum, Choi, Hyeongjoo, Jo, Yongsang, Kim, Bongsoo, Kang, Taejoon, Jung, Yongwon
Format: Article
Language:English
Subjects:
Avidin
Binding Sites
Biomolecules
Biosensors
DNA probes
miRNA
Models, Molecular
Molecular Probes - chemistry
multivalent interaction
Probes
Protein structure
Proteins
Ribonucleic acid
RNA
RNA - chemistry
RNA probes
RNA recognition
RNA-binding protein
RNA-Binding Proteins - chemistry
Scaffolds
SERS
specific interaction
Spectrum Analysis, Raman
Structural stability
Surface Properties
Valency
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Summary:By greatly enhancing binding affinities against target biomolecules, multivalent interactions provide an attractive strategy for biosensing. However, there is also a major concern for increased binding to nonspecific targets by multivalent binding. A range of charge‐engineered probes of a structure‐specific RNA binding protein PAZ as well as multivalent forms of these PAZ probes were constructed by using diverse multivalent avidin proteins (2‐mer, 4‐mer, and 24‐mer). Increased valency vastly enhanced the binding stability of PAZ to structured target RNA. Surprisingly, nonspecific RNA binding of multivalent PAZ can be reduced even below that of the PAZ monomer by controlling negative charges on both PAZ and multivalent avidin scaffolds. The optimized 24‐meric PAZ showed nearly irreversible binding to target RNA with negligible binding to nonspecific RNA, and this ultra‐specific 24‐meric PAZ probe allowed SERS detection of intact microRNAs at an attomolar level. Multivalent probe for a biosensor: An RNA binding monomer and multivalent scaffold proteins were optimized to develop a multivalent probe with near irreversible binding to target RNA and negligible binding to nonspecific RNA. The resulting ultra‐specific 24‐meric RNA binder allowed SERS detection of intact miRNAs at an attomolar level.
ISSN:1433-7851
1521-3773
DOI:10.1002/anie.201709153