Denatured mammalian protein mixtures exhibit unusually high solubility in nucleic acid-free pure water

Junichiro Futami, Haruna Fujiyama, Rie Kinoshita, Hidenori Nonomura, Tomoko Honjo, Hiroko Tada, Hirokazu Matsushita, Yoshito Abe, Kazuhiro Kakimi

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

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.

Original languageEnglish
Article numbere113295
JournalPloS one
Volume9
Issue number11
DOIs
Publication statusPublished - Nov 18 2014

Fingerprint

Solubility
Nucleic Acids
nucleic acids
solubility
Water
Proteins
proteins
water
Protein Unfolding
Intrinsically Disordered Proteins
Agglomeration
Biotechnology
Chloroform
Phenol
Eukaryota
Denaturation
Dialysis
thiocyanates
protein aggregates
cells

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Futami, J., Fujiyama, H., Kinoshita, R., Nonomura, H., Honjo, T., Tada, H., ... Kakimi, K. (2014). Denatured mammalian protein mixtures exhibit unusually high solubility in nucleic acid-free pure water. PloS one, 9(11), [e113295]. https://doi.org/10.1371/journal.pone.0113295

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.

In: PloS one, Vol. 9, No. 11, e113295, 18.11.2014.

Research output: Contribution to journalArticle

Futami, J, Fujiyama, H, Kinoshita, R, Nonomura, H, Honjo, T, Tada, H, Matsushita, H, Abe, Y & Kakimi, K 2014, 'Denatured mammalian protein mixtures exhibit unusually high solubility in nucleic acid-free pure water', PloS one, vol. 9, no. 11, e113295. https://doi.org/10.1371/journal.pone.0113295
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. In: PloS one. 2014 ; Vol. 9, No. 11.
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