Tighter Ligand Binding Can Compensate for Impaired Stability of an RNA-Binding Protein

It has been widely shown that ligand-binding residues, by virtue of their orientation, charge, and solvent exposure, often have a net destabilizing effect on proteins that is offset by stability conferring residues elsewhere in the protein. This structure-function trade-off can constrain possible ad...

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Main Authors: Wallis, C.P., Richman, T.R., Filipovska, A., Rackham, Oliver
Format: Journal Article
Language:English
Published: AMER CHEMICAL SOC 2018
Subjects:
Online Access:http://purl.org/au-research/grants/arc/DP140104111
http://hdl.handle.net/20.500.11937/91509
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author Wallis, C.P.
Richman, T.R.
Filipovska, A.
Rackham, Oliver
author_facet Wallis, C.P.
Richman, T.R.
Filipovska, A.
Rackham, Oliver
author_sort Wallis, C.P.
building Curtin Institutional Repository
collection Online Access
description It has been widely shown that ligand-binding residues, by virtue of their orientation, charge, and solvent exposure, often have a net destabilizing effect on proteins that is offset by stability conferring residues elsewhere in the protein. This structure-function trade-off can constrain possible adaptive evolutionary changes of function and may hamper protein engineering efforts to design proteins with new functions. Here, we present evidence from a large randomized mutant library screen that, in the case of PUF RNA-binding proteins, this structural relationship may be inverted and that active-site mutations that increase protein activity are also able to compensate for impaired stability. We show that certain mutations in RNA-protein binding residues are not necessarily destabilizing and that increased ligand-binding can rescue an insoluble, unstable PUF protein. We hypothesize that these mutations restabilize the protein via thermodynamic coupling of protein folding and RNA binding. ©
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spelling curtin-20.500.11937-915092023-05-04T06:35:42Z Tighter Ligand Binding Can Compensate for Impaired Stability of an RNA-Binding Protein Wallis, C.P. Richman, T.R. Filipovska, A. Rackham, Oliver Science & Technology Life Sciences & Biomedicine Biochemistry & Molecular Biology 23S RIBOSOMAL-RNA DELETERIOUS MUTATIONS ANTIBIOTIC-RESISTANCE MODULAR RECOGNITION FUNCTION TRADEOFFS STRUCTURAL BASES FOLDING ACTIVITY NUCLEIC-ACIDS DOMAIN-V EVOLUTION Catalytic Domain Humans Ligands Mutagenesis, Site-Directed Mutation Protein Binding Protein Folding Protein Stability RNA RNA-Binding Proteins Humans RNA-Binding Proteins RNA Ligands Mutagenesis, Site-Directed Catalytic Domain Protein Binding Protein Folding Mutation Protein Stability It has been widely shown that ligand-binding residues, by virtue of their orientation, charge, and solvent exposure, often have a net destabilizing effect on proteins that is offset by stability conferring residues elsewhere in the protein. This structure-function trade-off can constrain possible adaptive evolutionary changes of function and may hamper protein engineering efforts to design proteins with new functions. Here, we present evidence from a large randomized mutant library screen that, in the case of PUF RNA-binding proteins, this structural relationship may be inverted and that active-site mutations that increase protein activity are also able to compensate for impaired stability. We show that certain mutations in RNA-protein binding residues are not necessarily destabilizing and that increased ligand-binding can rescue an insoluble, unstable PUF protein. We hypothesize that these mutations restabilize the protein via thermodynamic coupling of protein folding and RNA binding. © 2018 Journal Article http://hdl.handle.net/20.500.11937/91509 10.1021/acschembio.8b00424 English http://purl.org/au-research/grants/arc/DP140104111 http://purl.org/au-research/grants/arc/DP170103000 http://purl.org/au-research/grants/arc/DP180101656 AMER CHEMICAL SOC fulltext
spellingShingle Science & Technology
Life Sciences & Biomedicine
Biochemistry & Molecular Biology
23S RIBOSOMAL-RNA
DELETERIOUS MUTATIONS
ANTIBIOTIC-RESISTANCE
MODULAR RECOGNITION
FUNCTION TRADEOFFS
STRUCTURAL BASES
FOLDING ACTIVITY
NUCLEIC-ACIDS
DOMAIN-V
EVOLUTION
Catalytic Domain
Humans
Ligands
Mutagenesis, Site-Directed
Mutation
Protein Binding
Protein Folding
Protein Stability
RNA
RNA-Binding Proteins
Humans
RNA-Binding Proteins
RNA
Ligands
Mutagenesis, Site-Directed
Catalytic Domain
Protein Binding
Protein Folding
Mutation
Protein Stability
Wallis, C.P.
Richman, T.R.
Filipovska, A.
Rackham, Oliver
Tighter Ligand Binding Can Compensate for Impaired Stability of an RNA-Binding Protein
title Tighter Ligand Binding Can Compensate for Impaired Stability of an RNA-Binding Protein
title_full Tighter Ligand Binding Can Compensate for Impaired Stability of an RNA-Binding Protein
title_fullStr Tighter Ligand Binding Can Compensate for Impaired Stability of an RNA-Binding Protein
title_full_unstemmed Tighter Ligand Binding Can Compensate for Impaired Stability of an RNA-Binding Protein
title_short Tighter Ligand Binding Can Compensate for Impaired Stability of an RNA-Binding Protein
title_sort tighter ligand binding can compensate for impaired stability of an rna-binding protein
topic Science & Technology
Life Sciences & Biomedicine
Biochemistry & Molecular Biology
23S RIBOSOMAL-RNA
DELETERIOUS MUTATIONS
ANTIBIOTIC-RESISTANCE
MODULAR RECOGNITION
FUNCTION TRADEOFFS
STRUCTURAL BASES
FOLDING ACTIVITY
NUCLEIC-ACIDS
DOMAIN-V
EVOLUTION
Catalytic Domain
Humans
Ligands
Mutagenesis, Site-Directed
Mutation
Protein Binding
Protein Folding
Protein Stability
RNA
RNA-Binding Proteins
Humans
RNA-Binding Proteins
RNA
Ligands
Mutagenesis, Site-Directed
Catalytic Domain
Protein Binding
Protein Folding
Mutation
Protein Stability
url http://purl.org/au-research/grants/arc/DP140104111
http://purl.org/au-research/grants/arc/DP140104111
http://purl.org/au-research/grants/arc/DP140104111
http://hdl.handle.net/20.500.11937/91509