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Intrinsic Disorder-Based Design of Stable Globular Proteins

Directed stabilization of globular proteins via substitution of a minimal number of amino acid residues is one of the most complicated experimental tasks. This work summarizes our research on the effect of amino acid substitutions on the protein stability utilizing the outputs of the analysis of int...

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Autores principales: Nagibina, Galina S., Glukhova, Ksenia A., Uversky, Vladimir N., Melnik, Tatiana N., Melnik, Bogdan S.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7022990/
https://www.ncbi.nlm.nih.gov/pubmed/31906016
http://dx.doi.org/10.3390/biom10010064
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author Nagibina, Galina S.
Glukhova, Ksenia A.
Uversky, Vladimir N.
Melnik, Tatiana N.
Melnik, Bogdan S.
author_facet Nagibina, Galina S.
Glukhova, Ksenia A.
Uversky, Vladimir N.
Melnik, Tatiana N.
Melnik, Bogdan S.
author_sort Nagibina, Galina S.
collection PubMed
description Directed stabilization of globular proteins via substitution of a minimal number of amino acid residues is one of the most complicated experimental tasks. This work summarizes our research on the effect of amino acid substitutions on the protein stability utilizing the outputs of the analysis of intrinsic disorder predisposition of target proteins. This allowed us to formulate the basis of one of the possible approaches to the stabilization of globular proteins. The idea is quite simple. To stabilize a protein as a whole, one needs to find its "weakest spot" and stabilize it, but the question is how this weak spot can be found in a query protein. Our approach is based on the utilization of the computational tools for the per-residue evaluation of intrinsic disorder predisposition to search for the "weakest spot" of a query protein (i.e., the region(s) with the highest local predisposition for intrinsic disorder). When such "weakest spot" is found, it can be stabilized through a limited number of point mutations by introducing order-promoting residues at hot spots, thereby increasing structural stability of a protein as a whole. Using this approach, we were able to obtain stable mutant forms of several globular proteins, such as Gαo, GFP, ribosome protein L1, and circular permutant of apical domain of GroEL.
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spelling pubmed-70229902020-03-12 Intrinsic Disorder-Based Design of Stable Globular Proteins Nagibina, Galina S. Glukhova, Ksenia A. Uversky, Vladimir N. Melnik, Tatiana N. Melnik, Bogdan S. Biomolecules Article Directed stabilization of globular proteins via substitution of a minimal number of amino acid residues is one of the most complicated experimental tasks. This work summarizes our research on the effect of amino acid substitutions on the protein stability utilizing the outputs of the analysis of intrinsic disorder predisposition of target proteins. This allowed us to formulate the basis of one of the possible approaches to the stabilization of globular proteins. The idea is quite simple. To stabilize a protein as a whole, one needs to find its "weakest spot" and stabilize it, but the question is how this weak spot can be found in a query protein. Our approach is based on the utilization of the computational tools for the per-residue evaluation of intrinsic disorder predisposition to search for the "weakest spot" of a query protein (i.e., the region(s) with the highest local predisposition for intrinsic disorder). When such "weakest spot" is found, it can be stabilized through a limited number of point mutations by introducing order-promoting residues at hot spots, thereby increasing structural stability of a protein as a whole. Using this approach, we were able to obtain stable mutant forms of several globular proteins, such as Gαo, GFP, ribosome protein L1, and circular permutant of apical domain of GroEL. MDPI 2019-12-30 /pmc/articles/PMC7022990/ /pubmed/31906016 http://dx.doi.org/10.3390/biom10010064 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Nagibina, Galina S.
Glukhova, Ksenia A.
Uversky, Vladimir N.
Melnik, Tatiana N.
Melnik, Bogdan S.
Intrinsic Disorder-Based Design of Stable Globular Proteins
title Intrinsic Disorder-Based Design of Stable Globular Proteins
title_full Intrinsic Disorder-Based Design of Stable Globular Proteins
title_fullStr Intrinsic Disorder-Based Design of Stable Globular Proteins
title_full_unstemmed Intrinsic Disorder-Based Design of Stable Globular Proteins
title_short Intrinsic Disorder-Based Design of Stable Globular Proteins
title_sort intrinsic disorder-based design of stable globular proteins
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7022990/
https://www.ncbi.nlm.nih.gov/pubmed/31906016
http://dx.doi.org/10.3390/biom10010064
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