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. 1993 Apr;64(4):1178–1186. doi: 10.1016/S0006-3495(93)81483-8

Intermolecular protein interactions in solutions of bovine lens beta L-crystallin. Results from 1/T1 nuclear magnetic relaxation dispersion profiles.

S H Koenig 1, R D Brown 3rd 1, A K Kenworthy 1, A D Magid 1, R Ugolini 1
PMCID: PMC1262435  PMID: 8388267

Abstract

We report the magnetic field dependence of 1/T1 of solvent water protons and deuterons (nuclear magnetic relaxation dispersion, or NMRD, profiles) for solutions of steer lens beta L-crystallin. Such data allow the study of intermolecular protein interactions over a wide concentration range, here 1-34% vol/vol, by providing a measure of the rotational relaxation time of solute macromolecules. We conclude that, for approximately less than 5% protein, the solute particles are noncompact, with a rotationally averaged volume approximately three times that of a compact 60-kD sphere. (Earlier results for alpha-crystallin, approximately 1,000 kD, from optical and osmotic measurements (Vérétout and Tardieu, 1989. J. Mol. Biol. 205:713-728), show a similar, approximately twofold, effect). At intermediate concentrations, to approximately 20% protein, there is evidence for limited association or oligomerization, as found for the structurally related gamma II-crystallin (Koenig et al. 1990. Biophys. J. 57:461-469), to a limiting size about two-thirds that of alpha-crystallin. The difference in NMRD behavior of the three classes of crystallins is consonant with their differing osmotic properties (Vérétout and Tardieu. J. Mol. Biol. 1989, 205:713-728; Kenworthy, McIntosh, and Magid. Biophys. J. 1992. 61:A477; Tardieu et al. 1992. Eur. Biophys. J. 21:1-12). We indicate how the unusual structures and interactions of these three classes of proteins can be combined to optimize transparency and minimize colloid osmotic difficulties in eye lens.

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Selected References

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  1. Asselbergs F. A., Koopmans M., van Venrooij W. J., Bloemendal H. Improved resolution of calf lens beta-crystallins. Exp Eye Res. 1979 Feb;28(2):223–228. doi: 10.1016/0014-4835(79)90133-7. [DOI] [PubMed] [Google Scholar]
  2. Bax B., Lapatto R., Nalini V., Driessen H., Lindley P. F., Mahadevan D., Blundell T. L., Slingsby C. X-ray analysis of beta B2-crystallin and evolution of oligomeric lens proteins. Nature. 1990 Oct 25;347(6295):776–780. doi: 10.1038/347776a0. [DOI] [PubMed] [Google Scholar]
  3. Beaulieu C. F., Clark J. I., Brown R. D., 3rd, Spiller M., Koenig S. H. Relaxometry of calf lens homogenates, including cross-relaxation by crystallin NH groups. Magn Reson Med. 1988 Sep;8(1):45–57. doi: 10.1002/mrm.1910080106. [DOI] [PubMed] [Google Scholar]
  4. Berbers G. A., Boerman O. C., Bloemendal H., de Jong W. W. Primary gene products of bovine beta-crystallin and reassociation behavior of its aggregates. Eur J Biochem. 1982 Nov 15;128(2-3):495–502. doi: 10.1111/j.1432-1033.1982.tb06992.x. [DOI] [PubMed] [Google Scholar]
  5. Bloemendal H., Hendriks I., Body P. Beta H- and beta L-crystallins are populations of aggregates. Exp Eye Res. 1990 Jun;50(6):711–713. doi: 10.1016/0014-4835(90)90118-e. [DOI] [PubMed] [Google Scholar]
  6. Brown R. D., 3rd, Koenig S. H. 1/T1 rho and low-field 1/T1 of tissue water protons arise from magnetization transfer to macromolecular solid-state broadened lines. Magn Reson Med. 1992 Nov;28(1):145–152. doi: 10.1002/mrm.1910280115. [DOI] [PubMed] [Google Scholar]
  7. Chiou S. H., Azari P., Himmel M. E., Lin H. K., Chang W. P. Physicochemical characterization of beta-crystallins from bovine lenses: hydrodynamic and aggregation properties. J Protein Chem. 1989 Feb;8(1):19–32. doi: 10.1007/BF01025076. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Driessen H. P., Bax B., Slingsby C., Lindley P. F., Mahadevan D., Moss D. S., Tickle I. J. Structure of oligomeric beta B2-crystallin: an application of the T2 translation function to an asymmetric unit containing two dimers. Acta Crystallogr B. 1991 Dec 1;47(Pt 6):987–997. doi: 10.1107/s0108768191009163. [DOI] [PubMed] [Google Scholar]
  10. Hallenga K., Koenig S. H. Protein rotational relaxation as studied by solvent 1H and 2H magnetic relaxation. Biochemistry. 1976 Sep 21;15(19):4255–4264. doi: 10.1021/bi00664a019. [DOI] [PubMed] [Google Scholar]
  11. Koenig S. H., Beaulieu C. F., Brown R. D., 3rd, Spiller M. Oligomerization and conformation change in solutions of calf lens gamma II-crystallin. Results from 1/T1 nuclear magnetic relaxation dispersion profiles. Biophys J. 1990 Mar;57(3):461–469. doi: 10.1016/S0006-3495(90)82562-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Koenig S. H., Brown R. D., 3rd, Adams D., Emerson D., Harrison C. G. Magnetic field dependence of 1/T1 of protons in tissue. Invest Radiol. 1984 Mar-Apr;19(2):76–81. doi: 10.1097/00004424-198403000-00002. [DOI] [PubMed] [Google Scholar]
  13. Koenig S. H., Brown R. D., 3rd, Gibson J. F., Ward R. J., Peters T. J. Relaxometry of ferritin solutions and the influence of the Fe3+ core ions. Magn Reson Med. 1986 Oct;3(5):755–767. doi: 10.1002/mrm.1910030511. [DOI] [PubMed] [Google Scholar]
  14. Koenig S. H., Brown R. D., 3rd, Spiller M., Chakrabarti B., Pande A. Intermolecular protein interactions in solutions of calf lens alpha-crystallin. Results from 1/T1 nuclear magnetic relaxation dispersion profiles. Biophys J. 1992 Mar;61(3):776–785. doi: 10.1016/S0006-3495(92)81882-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Koenig S. H., Brown R. D., 3rd, Ugolini R. A unified view of relaxation in protein solutions and tissue, including hydration and magnetization transfer. Magn Reson Med. 1993 Jan;29(1):77–83. doi: 10.1002/mrm.1910290114. [DOI] [PubMed] [Google Scholar]
  16. Koenig S. H., Brown R. D., 3rd, Ugolini R. Magnetization transfer in cross-linked bovine serum albumin solutions at 200 MHz: a model for tissue. Magn Reson Med. 1993 Mar;29(3):311–316. doi: 10.1002/mrm.1910290306. [DOI] [PubMed] [Google Scholar]
  17. Koenig S. H., Bryant R. G., Hallenga K., Jacob G. S. Magnetic cross-relaxation among protons in protein solutions. Biochemistry. 1978 Oct 3;17(20):4348–4358. doi: 10.1021/bi00613a037. [DOI] [PubMed] [Google Scholar]
  18. Koenig S. H., Schillinger W. E. Nuclear magnetic relaxation dispersion in protein solutions. I. Apotransferrin. J Biol Chem. 1969 Jun 25;244(12):3283–3289. [PubMed] [Google Scholar]
  19. Koenig S. H. Theory of relaxation of mobile water protons induced by protein NH moieties, with application to rat heart muscle and calf lens homogenates. Biophys J. 1988 Jan;53(1):91–96. doi: 10.1016/S0006-3495(88)83069-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lapatto R., Nalini V., Bax B., Driessen H., Lindley P. F., Blundell T. L., Slingsby C. High resolution structure of an oligomeric eye lens beta-crystallin. Loops, arches, linkers and interfaces in beta B2 dimer compared to a monomeric gamma-crystallin. J Mol Biol. 1991 Dec 20;222(4):1067–1083. doi: 10.1016/0022-2836(91)90594-v. [DOI] [PubMed] [Google Scholar]
  21. Morgan C. F., Schleich T., Caines G. H., Farnsworth P. N. Elucidation of intermediate (mobile) and slow (solidlike) protein motions in bovine lens homogenates by carbon-13 NMR spectroscopy. Biochemistry. 1989 Jun 13;28(12):5065–5074. doi: 10.1021/bi00438a025. [DOI] [PubMed] [Google Scholar]
  22. Piatigorsky J., Wistow G. The recruitment of crystallins: new functions precede gene duplication. Science. 1991 May 24;252(5009):1078–1079. doi: 10.1126/science.252.5009.1078. [DOI] [PubMed] [Google Scholar]
  23. Pierscionek B., Augusteyn R. C. Protein distribution patterns in concentric layers from single bovine lenses: changes with development and ageing. Curr Eye Res. 1988 Jan;7(1):11–23. doi: 10.3109/02713688809047015. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. Slingsby C., Bateman O. A. Quaternary interactions in eye lens beta-crystallins: basic and acidic subunits of beta-crystallins favor heterologous association. Biochemistry. 1990 Jul 17;29(28):6592–6599. doi: 10.1021/bi00480a007. [DOI] [PubMed] [Google Scholar]
  26. Slingsby C., Simpson A., Ferszt A., Bateman O. A., Nalini V. Molecular interactions in the eye lens. Biochem Soc Trans. 1991 Nov;19(4):853–858. doi: 10.1042/bst0190853. [DOI] [PubMed] [Google Scholar]
  27. Tardieu A., Vérétout F., Krop B., Slingsby C. Protein interactions in the calf eye lens: interactions between beta-crystallins are repulsive whereas in gamma-crystallins they are attractive. Eur Biophys J. 1992;21(1):1–12. doi: 10.1007/BF00195438. [DOI] [PubMed] [Google Scholar]
  28. Vérétout F., Delaye M., Tardieu A. Molecular basis of eye lens transparency. Osmotic pressure and X-ray analysis of alpha-crystallin solutions. J Mol Biol. 1989 Feb 20;205(4):713–728. doi: 10.1016/0022-2836(89)90316-1. [DOI] [PubMed] [Google Scholar]
  29. Vérétout F., Tardieu A. The protein concentration gradient within eye lens might originate from constant osmotic pressure coupled to differential interactive properties of crystallins. Eur Biophys J. 1989;17(2):61–68. doi: 10.1007/BF00257103. [DOI] [PubMed] [Google Scholar]
  30. Winter F., Kimmich R. NMR field-cycling relaxation spectroscopy of bovine serum albumin, muscle tissue, Micrococcus luteus and yeast. 14N1H-quadrupole dips. Biochim Biophys Acta. 1982 Nov 24;719(2):292–298. doi: 10.1016/0304-4165(82)90101-5. [DOI] [PubMed] [Google Scholar]
  31. Wistow G. J., Piatigorsky J. Lens crystallins: the evolution and expression of proteins for a highly specialized tissue. Annu Rev Biochem. 1988;57:479–504. doi: 10.1146/annurev.bi.57.070188.002403. [DOI] [PubMed] [Google Scholar]
  32. Zigler J. S., Jr Age-related changes in the polypeptide composition of beta-crystallin from bovine lens. Exp Eye Res. 1978 May;26(5):537–546. doi: 10.1016/0014-4835(78)90063-5. [DOI] [PubMed] [Google Scholar]
  33. Zigler J. S., Jr, Sidbury J. B., Jr Structure of calf lens beta-crystallins. Exp Eye Res. 1973 Jul;16(3):207–214. doi: 10.1016/0014-4835(73)90215-7. [DOI] [PubMed] [Google Scholar]

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