Denatured Mammalian Protein Mixtures Exhibit Unusually High Solubility in Nucleic Acid-Free Pure Water
Preventing protein aggregation is a major goal of biotechnology. Since protein aggregates are mainly comprised of unfolded proteins, protecting against denaturation is likely to assist solubility in an aqueous medium. Contrary to this concept, we found denatured total cellular protein mixture from m...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4236158/ https://www.ncbi.nlm.nih.gov/pubmed/25405999 http://dx.doi.org/10.1371/journal.pone.0113295 |
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author | Futami, Junichiro Fujiyama, Haruna Kinoshita, Rie Nonomura, Hidenori Honjo, Tomoko Tada, Hiroko Matsushita, Hirokazu Abe, Yoshito Kakimi, Kazuhiro |
author_facet | Futami, Junichiro Fujiyama, Haruna Kinoshita, Rie Nonomura, Hidenori Honjo, Tomoko Tada, Hiroko Matsushita, Hirokazu Abe, Yoshito Kakimi, Kazuhiro |
author_sort | Futami, Junichiro |
collection | PubMed |
description | Preventing protein aggregation is a major goal of biotechnology. Since protein aggregates are mainly comprised of unfolded proteins, protecting against denaturation is likely to assist solubility in an aqueous medium. Contrary to this concept, we found denatured total cellular protein mixture from mammalian cell kept high solubility in pure water when the mixture was nucleic acids free. The lysates were prepared from total cellular protein pellet extracted by using guanidinium thiocyanate-phenol-chloroform mixture of TRIzol, denatured and reduced total protein mixtures remained soluble after extensive dialysis against pure water. The total cell protein lysates contained fully disordered proteins that readily formed large aggregates upon contact with nucleic acids or salts. These findings suggested that the highly flexible mixtures of disordered proteins, which have fully ionized side chains, are protected against aggregation. Interestingly, this unusual solubility is characteristic of protein mixtures from higher eukaryotes, whereas most prokaryotic protein mixtures were aggregated under identical conditions. This unusual solubility of unfolded protein mixtures could have implications for the study of intrinsically disordered proteins in a variety of cells. |
format | Online Article Text |
id | pubmed-4236158 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-42361582014-11-21 Denatured Mammalian Protein Mixtures Exhibit Unusually High Solubility in Nucleic Acid-Free Pure Water Futami, Junichiro Fujiyama, Haruna Kinoshita, Rie Nonomura, Hidenori Honjo, Tomoko Tada, Hiroko Matsushita, Hirokazu Abe, Yoshito Kakimi, Kazuhiro PLoS One Research Article Preventing protein aggregation is a major goal of biotechnology. Since protein aggregates are mainly comprised of unfolded proteins, protecting against denaturation is likely to assist solubility in an aqueous medium. Contrary to this concept, we found denatured total cellular protein mixture from mammalian cell kept high solubility in pure water when the mixture was nucleic acids free. The lysates were prepared from total cellular protein pellet extracted by using guanidinium thiocyanate-phenol-chloroform mixture of TRIzol, denatured and reduced total protein mixtures remained soluble after extensive dialysis against pure water. The total cell protein lysates contained fully disordered proteins that readily formed large aggregates upon contact with nucleic acids or salts. These findings suggested that the highly flexible mixtures of disordered proteins, which have fully ionized side chains, are protected against aggregation. Interestingly, this unusual solubility is characteristic of protein mixtures from higher eukaryotes, whereas most prokaryotic protein mixtures were aggregated under identical conditions. This unusual solubility of unfolded protein mixtures could have implications for the study of intrinsically disordered proteins in a variety of cells. Public Library of Science 2014-11-18 /pmc/articles/PMC4236158/ /pubmed/25405999 http://dx.doi.org/10.1371/journal.pone.0113295 Text en © 2014 Futami et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Futami, Junichiro Fujiyama, Haruna Kinoshita, Rie Nonomura, Hidenori Honjo, Tomoko Tada, Hiroko Matsushita, Hirokazu Abe, Yoshito Kakimi, Kazuhiro Denatured Mammalian Protein Mixtures Exhibit Unusually High Solubility in Nucleic Acid-Free Pure Water |
title | Denatured Mammalian Protein Mixtures Exhibit Unusually High Solubility in Nucleic Acid-Free Pure Water |
title_full | Denatured Mammalian Protein Mixtures Exhibit Unusually High Solubility in Nucleic Acid-Free Pure Water |
title_fullStr | Denatured Mammalian Protein Mixtures Exhibit Unusually High Solubility in Nucleic Acid-Free Pure Water |
title_full_unstemmed | Denatured Mammalian Protein Mixtures Exhibit Unusually High Solubility in Nucleic Acid-Free Pure Water |
title_short | Denatured Mammalian Protein Mixtures Exhibit Unusually High Solubility in Nucleic Acid-Free Pure Water |
title_sort | denatured mammalian protein mixtures exhibit unusually high solubility in nucleic acid-free pure water |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4236158/ https://www.ncbi.nlm.nih.gov/pubmed/25405999 http://dx.doi.org/10.1371/journal.pone.0113295 |
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