Skip to main content
SpringerLink
Log in

Protein interactions in the calf eye lens: interactions between β-crystallins are repulsive whereas in γ-crystallins they are attractive

  • Published:
European Biophysics Journal Aims and scope Submit manuscript

Abstract

Non-specific interactions in β- and γ-crystallins have been studied by solution X-ray scattering and osmotic pressure experiments. Measurements were carried out as a function of protein concentration at two ionic strengths. The effect of temperature was tested between 7°C and 31°C. Two types of interactions were observed. With β-crystallin solutions, a repulsive coulombic interaction could be inferred from the decrease of the normalized X-ray scattering intensity near the origin with increasing protein concentration and from the fact that the osmotic pressure increases much more rapidly than in the ideal case. As was previously observed with α-crystallins, such behaviour is dependent upon ionic strength but is hardly affected by temperature. In contrast, with γ-crystallin solutions, the normalized X-ray scattering intensity near the origin increases with increasing protein concentration and the osmotic pressure increases less rapidly than in the ideal case. Such behaviour indicates that attractive forces are predominant, although we do not yet know their molecular origin. Under our experimental conditions, the effect of temperature was striking whereas no obvious contribution of the ionic strength could be seen, perhaps owing to masking by the large temperature effect. The relevance of the different types of non-specific interactions for lens function is discussed.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Aarts HJM, Lubsen NH, Schoenmakers JGG (1989) Crystallin gene expression during rat lens development. Eur J Biochem 183:31–36

    Google Scholar 

  • Asselbergs FAM, Koopmans M, van Venrooij WJ, Bloemendal H (1979) Improved resolution of calf lens β-crystallins. Exp Eye Res 28:223–228

    Google Scholar 

  • Barlow DJ, Thornton JM (1983) Ion pairs in proteins. J Mol Biol 168:867–885

    Google Scholar 

  • Bax B, Lapatto R, Nalini V, Driessen H, Lindley PF, Mahadevan D, Blundell TL, Slingsby C (1990) X-ray analysis of βB2-crystallin and evolution of oligomeric lens proteins. Nature 347:776–780

    Google Scholar 

  • Belloni L (1986) Electrostatic interactions in colloidal solutions: comparison between primitive and one-component models. J Chem Phys 85:519–526

    Google Scholar 

  • Benedek GG (1971) Theory of transparency of the eye. Appl Opt 10:459–473

    Google Scholar 

  • Berbers GAM, Boermann OC, Bloemendal H, de Jong WW (1982) Primary gene products of bovine β-crystallin and reassociation behaviour of its aggregates. Eur J Biochem 128:495–502

    Google Scholar 

  • Berbers GAM, Hoekman WA, Bloemendal H, de Jong WW, Kleinschmidt T, Braunitzer G (1984) Homology between the primary structures of the major β-crystallin chains. Eur J Biochem 139:467–479

    Google Scholar 

  • Bindels JG, Koppers A, Hoenders HJ (1981) Structural aspects of bovine β-crystallins: physical characterization including dissociation-association behaviour. Exp Eye Res 33:333–343

    Google Scholar 

  • Björk I (1961) Studies on γ-crystallin from calf lens: Isolation by gel filtration. Exp Eye Res 1:145–154

    Google Scholar 

  • Björk I (1964) Studies on γ-crystallin from calf lens II. Purification and some properties of the main protein components. Exp Eye Res 3:254–258

    Google Scholar 

  • Bloemendal H, Piatigorsky J, Spector A (1989) Recommendations for crystallin nomenclature. Exp Eye Res 48:465–466

    Google Scholar 

  • Blundell TL, Lindley PF, Miller L, Moss DS, Slingsby C, Tickle IJ, Turnell WG, Wistow GJ (1981 a) The molecular structure and stability of the eye lens: X-ray analysis of γ-crystallin II. Nature 289:771–777

    Google Scholar 

  • Blundell TL, Lindley PF, Miller L, Moss DS, Slingsby C, Tickle IJ, Turnell WG, Wistow GJ (1981 b) The molecular structure of γ-crystallin in relation to eye lens transparency. Lens Res 1:109–131

    Google Scholar 

  • Blundell TL, Lindley PF, Miller LR, Moss DS, Slingsby C, Turnell WG, Wistow GJ (1983) Interactions of γ-crystallins in relation to eye-lens transparency. Lens Res 1:109–131

    Google Scholar 

  • Bordas J, Koch MHJ, Clout PN, Dorrington E, Boulin C, Gabriel A (1980) A synchrotron radiation camera and data acquisition system for time resolved X-ray scattering studies. J Phys Sci Instrum E13:938–944

    Google Scholar 

  • Broide ML, Berland CR, Pande J, Ogun OO, Benedek GB (1991) Binary liquid phase separation of lens protein solutions. PNAS 88:5660–5664

    Google Scholar 

  • Chirgadze YN, Nevskaya NA, Fomenkova NP, Niklonov SV, Sergeev YV, Brazhnikov EV, Garber MB, Lunin VY, Urzumtsev AP, Vernoslova EA (1986) Spatial structure of γ-crystallin IIIb from calf eye lens at 25 Å resolution. Dokl Akad Nauk USSR 290:492–495

    Google Scholar 

  • Clark JI, Benedek GB (1980) Phase diagram for cell cytoplasm from the calf lens. Biochem Biophys Res Commun 95:482–489

    Google Scholar 

  • Clark JI, Carper D (1987) Phase separation in lens cytoplasm is genetically linked to cataract formation in the Philly mouse. PNAS 84:122–125

    Google Scholar 

  • Cother E, Kwan B, Beaty C (1968) The lens as an osmometer and the effects of medium osmolarity on water transport, 86 Rb efflux and 86 Rb transport by the lens. Biochim Biophys Acta 150:705–722

    Google Scholar 

  • De Jong WW, Hendriks W, Mulders JWM, Bloemendal H (1989) Evolution of eye lens crystallins: the stress connection. Trends Biochem Sci 14:365–368

    Google Scholar 

  • Delaye M, Tardieu A (1983) Short range order of crystallin proteins account for eye lens transparency. Nature 302:415–417

    Google Scholar 

  • Delaye M, Clark JI, Benedek GB (1981) Coexistence curve for phase separation in the lens cytoplasm. Biochem Biophys Res Commun 100:908–914

    Google Scholar 

  • Delaye M, Clark JI, Benedek GB (1982) Identification of the scattering elements responsible for lens opacification in cold lens cataracts. Biophys J 37:647–656

    Google Scholar 

  • Delaye M, Laporte D, Clark J, Tardieu A, Gulik-Krzywicki T (1986) Model systems of lens opacification: cold cataract and calcium induced cataract. Modern Trends in aging research Colloque INSERM-EURAGE, John Libbey Eurotext Ldt 147:357–364

    Google Scholar 

  • Den Dunnen JT, Moormann RJM, Lubsen NH, Schoenmakers JGG (1986) Concerted and divergent evolution within the rat γ-gene family. J Mol Biol 189:37–46

    Google Scholar 

  • Depautex C, Desvignes C, Feder P, Lemonnier M, Bosshard R, Leboucher P, Dageaux D, Benoit JP, Vachette P (1987) Le banc de diffusion des rayons X aux petits angles D24. LURE Rapport d'activité pour la période Août 1985–1987, doc, CEN Saclay, 1987

  • Eisenberg H (1976) Biological macromolecules and polyelectrolytes in solution. Clarendon Press, Oxford

    Google Scholar 

  • Guinier A, Fournet G (1955) Small angle scattering of X-rays. Wiley, New York

    Google Scholar 

  • Gulik-Krzywicki T, Tardieu A, Delaye M (1984) Spatial reorganization of low molecular weight proteins during cold cataract opacification. Biochim Biophys Acta 800:28–32

    CAS  PubMed  Google Scholar 

  • Hayter JB, Penfold J (1981) An analytical structure factor for macroion solutions. Mol Phys 42:109–118

    Google Scholar 

  • Hansen JP, Hayter JB (1982) A rescaled MSA structure factor for dilute charged colloidal dispersions. Mol Phys 46:651–656

    Google Scholar 

  • Harding JJ, Crabbe MJC (1984) The lens: development, proteins, metabolism and cataract. In: Davson H (ed) The eye, vol 1B, 3rd edn. Academic Press, London, pp 207–492

    Google Scholar 

  • Huizinga A, Bot ACC, De Mul FFM, Vrensen GFJM, Greve J (1989) Local variation in absolute water content of human and rabbit lenses measured by Raman spectroscopy. Exp Eye Res 48:187–496

    Google Scholar 

  • Ingolia TD, Craig EA (1982) Four small Drosophila heat shock proteins are related to each other and to mammalian α-crystallin. PNAS 79:2360–2364

    Google Scholar 

  • Iwaki T, Kume-Iwaki A, Liem RKH, Goldman JE (1989) αB-crystallin is expressed in non-lenticular tissues and accumulates in Alexander's disease brain. Cell 57:71–78

    Google Scholar 

  • Lubsen NH, Aarts HJM, Schoenmakers JGG (1988) The evolution of lenticular proteins: the β- and the γ-gene super-family. Prog Biophys Mol Biol 51:47–76

    Google Scholar 

  • Luzzati V, Tardieu A (1980) Recent developments in solution X-ray scattering. Ann Rev Biophys Bioeng 9:1–29

    Google Scholar 

  • Ninham BW (1982) Hierarchies of forces: the last 150 years. Adv Coll Interface Sci 16:3–15

    Google Scholar 

  • Parsegian VA, Rand RP, Fuller NL, Rau DC (1986) Osmotic stress for the direct measurement of intermolecular forces. Methods Enzymol 127:400–416

    Article  CAS  PubMed  Google Scholar 

  • Piatigorsky J (1981) Lens differenciation in vertebrates. A review of cellular and molecular features. Differenciation 19:134–153

    Google Scholar 

  • Piatigorsky J (1984) Lens crystallin and their gene families. Cell 38:620–621

    Google Scholar 

  • Piatigorsky J, Wistow G (1989) Enzyme/crystallins: gene sharing as an evolutionary strategy. Cell 57:197–199

    Google Scholar 

  • Piatigorsky J, Wistow G (1991) The recruitment of crystallins: new functions precede gene duplication. Science 252:1078–1079

    CAS  PubMed  Google Scholar 

  • Prouty MS, Schechter AN, Parsegian VA (1985) Chemical potential measurements of Deoxyhemoglobin S Polymerisation Determination of the phase diagram of an assembling protein. J Mol Biol 184:517–528

    Google Scholar 

  • Reiff TR (1987) Colloid osmotic pressure of cortical and nuclear lenticular homogenates. Clin Res 35:98A

    Google Scholar 

  • Sergeev YV, Chirgadze YN, Mylvaganam SE, Driessen H, Slingsby C, Blundell TL (1988) Surface interactions of γ-crystallins in the crystal medium in relation to their association in the eye. Lens Proteins: Struct Funct Genet 4:137–147

    Google Scholar 

  • Siezen RJ, Benedek GB (1985) Controlled modulation of the phase separation and opacification temperature of purified bovine γIV-crystallin. Current Eye Res 4:1077–1085

    Google Scholar 

  • Siezen RJ, Fisch MR, Slingsby C, Benedek GB (1985) Opacification of γ-crystallin solutions from calf lens in relation to cold cataract formation. Proc Natl Acad Sci USA 82:1701–1705

    Google Scholar 

  • Siezen RJ, Anello RD, Thomson JA (1986) Interactions of lens proteins concentration dependence of β-crystallin aggregation. 43:293–303

    Google Scholar 

  • Siezen RJ, Thomson J, Kaplan ED, Benedek GB (1987) Human lens γ-crystallins: Isolation, identification and characterisation of the expressed gene products. Proc Natl Acad Sci USA 84:6088–6092

    Google Scholar 

  • Siezen RJ, Wu E, Kaplan ED, Thomson JA, Benedek GB (1988) Rat lens γ-crystallins: characterization of the six gene products and their spatial and temporal distribution resulting from differential synthesis. J Mol Biol 199:475–490

    Google Scholar 

  • Slingsby C (1985) Structural variation in lens crystallins. TIBS 10:281–284

    Google Scholar 

  • Slingsby C, Bateman OA (1990) Quaternary interactions in eye lens β-crystallins: basic and acidic subunits of β-crystallins favor heterologous association. Biochemistry 29:6592–6599

    Google Scholar 

  • Slingsby C, Miller L (1983) Purification and crystallisation of mammalian γ-crystallins. Exp Eye Res 37:517–530

    Google Scholar 

  • Slingsby C, Miller L (1985) The reaction of glutathione with eye-lens protein γ-crystallin. Biochem J 230:143–150

    Google Scholar 

  • Slingsby C, Driessen HPC, Mahadevan D, Bax B, Blundell TL (1988) Evolutionary and functional relationships between the basic and acidic γ-crystallins. Exp Eye Res 46:375–403

    Google Scholar 

  • Summers L, Wistow G, Narebor M, Moss D, Lindley P, Slingsby C, Blundell T, Bartunik H, Bartels K (1984) X-ray studies of the lens specific proteins: the crystallins. In: Hearn MTW (ed) Peptide and protein reviews, vol 3. Dekker, New York, pp 147–168

    Google Scholar 

  • Summers L, Slingsby C, Blundell TL, Den Dunnen JT, Moormann RJM, Schoenmakers JGG (1986) Structural variation in mammalian γ-crystallins based on computer graphics analyses of human, rat and calf sequences. 1-Core packing and surface properties. Exp Eye Res 43:77–92

    Google Scholar 

  • Tanaka T, Benedek GB (1975) Observation of protein diffusity in intact human and bovine lenses with application to cataract. Invest Ophthalmol 14:449–456

    Google Scholar 

  • Tanaka T, Ishimoto C, Chylack LT (1977) Phase separation of a protein-water mixture in cold cataract in the young rat lens. Science 197:1010–1012

    Google Scholar 

  • Tardieu A, Delaye M (1988) Eye lens proteins and transparency: from light transmission theory to solution X-ray structural analysis. Ann Rev Biophys Chem 17:1207–1215

    Google Scholar 

  • Tardieu A, Laporte D, Delaye M (1987) Colloidal dispersions of α-crystallin proteins. Small angle X-ray analysis of the dispersion structure. J Phys 48:1207–1209

    Google Scholar 

  • Thomson JA, Schurtenberger P, Thruston GM, Benedek GB (1987) Binary liquid phase separation and critical phenomena in a protein-water solution. Proc Natl Acad Sci USA 84:7079–7083

    Google Scholar 

  • Vérétout F, Tardieu A (1989) The protein concentration gradient within eye lens might originate from constant osmotic pressure coupled to differential interactive properties of crystallins. Eur Biophys J 17:61–68

    Google Scholar 

  • Vérétout F, Delaye M, Tardieu A (1989) Molecular basis of eye lens transparency: Osmotic pressure and X-ray analysis of alphacrystallin solutions. J Mol Biol 205:713–728

    Google Scholar 

  • Verwey EJW Overbeek JTG (1948) Theory of the stability of lyophobic colloids. Elsevier, Amsterdam

    Google Scholar 

  • Voorter CEM, De Haard-Hoekman WA, Hermans MMP, Bloemendal H, De Jong WW (1990) Differential synthesis of crystallins in the developing rat eye lens. Exp Eye Res 50:429–438

    Google Scholar 

  • Waley SG (1969) The lens: function and macromolecular composition. In: Davson H (ed) The eye, vol 1, 2nd edn. Academic Press, London, pp 299–379

    Google Scholar 

  • Wang X, Bettelheim FA (1989) Second virial coefficient of α-crystallin proteins. Struct Funct Genet 5:166–169

    Google Scholar 

  • White HE, Driessen HPC, Slingsby C, Moss DS, Lindley PF (1989) Packing interactions in the eye lens: Structural analysis, internal symmetry and lattice interactions of bovine γIVa-crystallin. J Mol Biol 207:217–235

    Google Scholar 

  • Wistow G, Piatigorsky J (1987) Recruitment of enzymes and the lens structural proteins. Science 236:1554–1556

    Google Scholar 

  • Wistow G, Piatigorsky J (1988) Lens crystallins: The evolution and the expression of protein for a highly specialized tissue. Ann Rev Biochem 57:479–504

    Google Scholar 

  • Wistow G, Slingsby C, Blundell T, Driessen H, De Jong W, Bloemendal H (1981) Eye-lens proteins: The three-dimensional structure of β-crystallin predicted from monomeric γ-crystallin. FEBS Lett 133:9–16

    Google Scholar 

  • Wistow G, Turnell B, Summers L, Slingsby C, Moss D, Miller L, Lindley P, Blundell T (1983) X-ray analysis of the eye lens protein γII-crystallin at 19 Å resolution. J Mol Biol 170:175–202

    Google Scholar 

  • Zigman S, Lerman S (1964) A cold precipitable protein in the lens. Nature 203:662–663

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Centre de Génétique Moléculaire, CNRS, F-91198, Gif sur Yvette Cedex, France

    Annette Tardieu, Françoise Vérétout & Brigitte Krop

  2. Laboratory of Molecular Biology, Department of Crystallography, Birkbeck College, Malet Street, WCIE 7HX, London, United Kingdom

    Christine Slingsby

Authors
  1. Annette Tardieu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Françoise Vérétout
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Brigitte Krop
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Christine Slingsby
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Offprint requests to: A. Tardieu

Rights and permissions

Reprints and permissions

About this article

Cite this article

Tardieu, A., Vérétout, F., Krop, B. et al. Protein interactions in the calf eye lens: interactions between β-crystallins are repulsive whereas in γ-crystallins they are attractive. Eur Biophys J 21, 1–12 (1992). https://doi.org/10.1007/BF00195438

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00195438

Key words

Search

Navigation