Skip to main content
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1987 Oct;84(20):7079–7083. doi: 10.1073/pnas.84.20.7079

Binary liquid phase separation and critical phenomena in a protein/water solution.

J A Thomson 1, P Schurtenberger 1, G M Thurston 1, G B Benedek 1
PMCID: PMC299233  PMID: 3478681

Abstract

We have investigated the phase diagram of aqueous solutions of the bovine lens protein gamma II-crystallin. For temperatures T less than Tc = 278.5 K, we find that these solutions exhibit a reversible coexistence between two isotropic liquid phases differing in protein concentration. The dilute and concentrated branches of the coexistence curve were characterized, consistently, both by measurements of the two coexisting concentrations, c(T), and by measuring the cloud temperatures for various initial concentrations. We estimate that the critical concentration, cc, is 244 mg of protein per ml solution. The coexistence curve is well represented by the absolute value of (c - cc)/cc = 5.2 square root (Tc - T)/Tc. Using the temperature dependence of the scattered light intensity along isochores parallel to the critical isochore, we estimated the location of the spinodal line and found it to have the form (c - cc)/cc = 3.0 square root (Tc - T)/Tc. The ratio of the widths of the coexistence curve and the spinodal line, (5.2/3.0), is close to the mean-field value square root 3. We have also observed the growth of large crystals of gamma II-crystallin in some of these aqueous solutions and have made preliminary observations as to the factors that promote or delay the onset of crystallization. These findings suggest that selected protein/water systems can serve as excellent model systems for the study of phase transitions and critical phenomena.

Full text

PDF
7083

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bhat S. P., Spector A. Complete nucleotide sequence of a cDNA derived from calf lens gamma-crystallin mRNA: presence of Alu I-like DNA sequences. DNA. 1984 Aug;3(4):287–295. doi: 10.1089/dna.1.1984.3.287. [DOI] [PubMed] [Google Scholar]
  2. Blundell T., Lindley P., Miller L., Moss D., Slingsby C., Tickle I., Turnell B., Wistow G. The molecular structure and stability of the eye lens: x-ray analysis of gamma-crystallin II. Nature. 1981 Feb 26;289(5800):771–777. doi: 10.1038/289771a0. [DOI] [PubMed] [Google Scholar]
  3. Brandau D. T., Trautman P. A., Steadman B. L., Lawson E. Q., Middaugh C. R. The temperature-dependent stoichiometry of mixed cryoimmunoglobulins. J Biol Chem. 1986 Dec 15;261(35):16385–16391. [PubMed] [Google Scholar]
  4. Clark J. I., Carper D. Phase separation in lens cytoplasm is genetically linked to cataract formation in the Philly mouse. Proc Natl Acad Sci U S A. 1987 Jan;84(1):122–125. doi: 10.1073/pnas.84.1.122. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Clark J. I., Giblin F. J., Reddy V. N., Benedek G. B. Phase separation of X-irradiated lenses of rabbit. Invest Ophthalmol Vis Sci. 1982 Feb;22(2):186–190. [PubMed] [Google Scholar]
  6. Croft L. R. The amino acid sequence of -crystallin (fraction II) from calf lens. Biochem J. 1972 Jul;128(4):961–970. doi: 10.1042/bj1280961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Ishimoto C., Goalwin P. W., Sun S. T., Nishio I., Tanaka T. Cytoplasmic phase separation in formation of galactosemic cataract in lenses of young rats. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4414–4416. doi: 10.1073/pnas.76.9.4414. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Middaugh C. R., Gerber-Jenson B., Hurvitz A., Paluszek A., Scheffel C., Litman G. W. Physicochemical characterization of six monoclonal cryoimmunoglobulins: possible basis for cold-dependent insolubility. Proc Natl Acad Sci U S A. 1978 Jul;75(7):3440–3444. doi: 10.1073/pnas.75.7.3440. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Noguchi C. T., Schechter A. N. Sickle hemoglobin polymerization in solution and in cells. Annu Rev Biophys Biophys Chem. 1985;14:239–263. doi: 10.1146/annurev.bb.14.060185.001323. [DOI] [PubMed] [Google Scholar]
  10. Phillies GD. Comment on "Critical behavior of a binary mixture of protein and salt water". Phys Rev Lett. 1985 Sep 16;55(12):1341–1341. doi: 10.1103/PhysRevLett.55.1341. [DOI] [PubMed] [Google Scholar]
  11. Prouty M. S., Schechter A. N., Parsegian V. A. Chemical potential measurements of deoxyhemoglobin S polymerization. Determination of the phase diagram of an assembling protein. J Mol Biol. 1985 Aug 5;184(3):517–528. doi: 10.1016/0022-2836(85)90298-0. [DOI] [PubMed] [Google Scholar]
  12. Schoenmakers J. G., den Dunnen J. T., Moormann R. J., Jongbloed R., van Leen R. W., Lubsen N. H. The crystallin gene families. Ciba Found Symp. 1984;106:208–218. doi: 10.1002/9780470720875.ch12. [DOI] [PubMed] [Google Scholar]
  13. Siezen R. J., Benedek G. B. Controlled modulation of the phase separation and opacification temperature of purified bovine gamma IV-crystallin. Curr Eye Res. 1985 Oct;4(10):1077–1085. doi: 10.3109/02713688509003352. [DOI] [PubMed] [Google Scholar]
  14. Siezen R. J., Fisch M. R., Slingsby C., Benedek G. B. Opacification of gamma-crystallin solutions from calf lens in relation to cold cataract formation. Proc Natl Acad Sci U S A. 1985 Mar;82(6):1701–1705. doi: 10.1073/pnas.82.6.1701. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Summers L., Slingsby C., White H., Narebor M., Moss D., Miller L., Mahadevan D., Lindley P., Driessen H., Blundell T. The molecular structures and interactions of bovine and human gamma-crystallins. Ciba Found Symp. 1984;106:219–236. doi: 10.1002/9780470720875.ch13. [DOI] [PubMed] [Google Scholar]
  16. Tanaka T., Benedek G. B. Observation of protein diffusivity in intact human and bovine lenses with application to cataract. Invest Ophthalmol. 1975 Jun;14(6):449–456. [PubMed] [Google Scholar]
  17. Thomson J. A., Augusteyn R. C. Ontogeny of human lens crystallins. Exp Eye Res. 1985 Mar;40(3):393–410. doi: 10.1016/0014-4835(85)90152-6. [DOI] [PubMed] [Google Scholar]
  18. Wistow G., Turnell B., Summers L., Slingsby C., Moss D., Miller L., Lindley P., Blundell T. X-ray analysis of the eye lens protein gamma-II crystallin at 1.9 A resolution. J Mol Biol. 1983 Oct 15;170(1):175–202. doi: 10.1016/s0022-2836(83)80232-0. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES