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. 2000 Aug 15;350(Pt 1):329–335.

The analysis of intermolecular interactions in concentrated hyaluronan solutions suggest no evidence for chain-chain association.

P Gribbon 1, B C Heng 1, T E Hardingham 1
PMCID: PMC1221259  PMID: 10926861

Abstract

Confocal fluorescence recovery after photobleaching (confocal-FRAP) was used to examine the influence of electrolytes (NaCl, KCl, MgCl(2), MnCl(2) and CaCl(2)) on the network and hydrodynamic properties of fluoresceinamine-labelled hyaluronan (FA-HA) at concentrations up to 10 mg/ml. Self and tracer lateral diffusion coefficients showed that in Ca(2+) and Mn(2+), FA-HA (830 kDa) was more compact than in Mg(2+), Na(+) or K(+). These results were correlated with changes in the hydrodynamic radius of HA, determined by multi-angle laser-light-scattering analysis in dilute solution, which was smaller in CaCl(2) (36 nm) than in NaCl (43 nm). The permeability of more concentrated solutions of HA (<10 mg/ml) to FITC-dextran tracers (2000 kDa) was higher in CaCl(2). The properties of HA in urea (up to 6 M) were investigated to test for hydrophobic interactions and also in ethanol/water (up to 62%, v/v). In both, there was reduced hydrodynamic size and increased permeability to FITC-dextran, suggesting increased chain flexibility, but it did not show the changes predicted if chain-chain association was disrupted by urea, or enhanced by ethanol. Oligosaccharides of HA (HA(20-26)) also had no effect on the self diffusion of high-molecular-mass FA-HA (830 kDa) solutions, or on dextran tracer diffusion, showing that there were no chain-chain interactions open to competition by short-chain segments. The results suggest that the effects of electrolytes and solvent are determined primarily by their effect on HA chain flexibility, with no evidence for association between chain segments contributing significantly to the major properties.

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Selected References

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  1. Almond A., Brass A., Sheehan J. K. Deducing polymeric structure from aqueous molecular dynamics simulations of oligosaccharides: predictions from simulations of hyaluronan tetrasaccharides compared with hydrodynamic and X-ray fibre diffraction data. J Mol Biol. 1998 Dec 18;284(5):1425–1437. doi: 10.1006/jmbi.1998.2245. [DOI] [PubMed] [Google Scholar]
  2. Almond A., Brass A., Sheehan J. K. Dynamic exchange between stabilized conformations predicted for hyaluronan tetrasaccharides: comparison of molecular dynamics simulations with available NMR data. Glycobiology. 1998 Oct;8(10):973–980. doi: 10.1093/glycob/8.10.973. [DOI] [PubMed] [Google Scholar]
  3. Almond A., Sheehan J. K., Brass A. Molecular dynamics simulations of the two disaccharides of hyaluronan in aqueous solution. Glycobiology. 1997 Jul;7(5):597–604. doi: 10.1093/glycob/7.5.597. [DOI] [PubMed] [Google Scholar]
  4. Cowman M. K., Cozart D., Nakanishi K., Balazs E. A. 1H NMR of glycosaminoglycans and hyaluronic acid oligosaccharides in aqueous solution: the amide proton environment. Arch Biochem Biophys. 1984 Apr;230(1):203–212. doi: 10.1016/0003-9861(84)90101-2. [DOI] [PubMed] [Google Scholar]
  5. Fujii K., Kawata M., Kobayashi Y., Okamoto A., Nishinari K. Effects of the addition of hyaluronate segments with different chain lengths on the viscoelasticity of hyaluronic acid solutions. Biopolymers. 1996 May;38(5):583–591. doi: 10.1002/(SICI)1097-0282(199605)38:5%3C583::AID-BIP4%3E3.0.CO;2-O. [DOI] [PubMed] [Google Scholar]
  6. Glabe C. G., Harty P. K., Rosen S. D. Preparation and properties of fluorescent polysaccharides. Anal Biochem. 1983 Apr 15;130(2):287–294. doi: 10.1016/0003-2697(83)90590-0. [DOI] [PubMed] [Google Scholar]
  7. Gribbon P., Hardingham T. E. Macromolecular diffusion of biological polymers measured by confocal fluorescence recovery after photobleaching. Biophys J. 1998 Aug;75(2):1032–1039. doi: 10.1016/S0006-3495(98)77592-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gribbon P., Heng B. C., Hardingham T. E. The molecular basis of the solution properties of hyaluronan investigated by confocal fluorescence recovery after photobleaching. Biophys J. 1999 Oct;77(4):2210–2216. doi: 10.1016/S0006-3495(99)77061-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Lapcík L Jr and Lubomír, Lapcík Lubomír, De Smedt Stefaan, Demeester Joseph, Chabrecek Peter. Hyaluronan: Preparation, Structure, Properties, and Applications. Chem Rev. 1998 Dec 17;98(8):2663–2684. doi: 10.1021/cr941199z. [DOI] [PubMed] [Google Scholar]
  10. Maroudas A., Weinberg P. D., Parker K. H., Winlove C. P. The distributions and diffusivities of small ions in chondroitin sulphate, hyaluronate and some proteoglycan solutions. Biophys Chem. 1988 Dec;32(2-3):257–270. doi: 10.1016/0301-4622(88)87012-1. [DOI] [PubMed] [Google Scholar]
  11. Morris E. R., Rees D. A., Welsh E. J. Conformation and dynamic interactions in hyaluronate solutions. J Mol Biol. 1980 Apr;138(2):383–400. doi: 10.1016/0022-2836(80)90294-6. [DOI] [PubMed] [Google Scholar]
  12. Parker K. H., Winlove C. P., Maroudas A. The theoretical distributions and diffusivities of small ions in chondroitin sulphate and hyaluronate. Biophys Chem. 1988 Dec;32(2-3):271–282. doi: 10.1016/0301-4622(88)87013-3. [DOI] [PubMed] [Google Scholar]
  13. Reed C. E., Li X., Reed W. F. The effects of pH on hyaluronate as observed by light scattering. Biopolymers. 1989 Nov;28(11):1981–2000. doi: 10.1002/bip.360281114. [DOI] [PubMed] [Google Scholar]
  14. Scott J. E., Cummings C., Brass A., Chen Y. Secondary and tertiary structures of hyaluronan in aqueous solution, investigated by rotary shadowing-electron microscopy and computer simulation. Hyaluronan is a very efficient network-forming polymer. Biochem J. 1991 Mar 15;274(Pt 3):699–705. doi: 10.1042/bj2740699. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Scott J. E., Heatley F. Hyaluronan forms specific stable tertiary structures in aqueous solution: a 13C NMR study. Proc Natl Acad Sci U S A. 1999 Apr 27;96(9):4850–4855. doi: 10.1073/pnas.96.9.4850. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Scott J. E., Tigwell M. J. The influence of the intrapolymer environment on periodate oxidation of uronic acids in polyuronides and glycosaminoglycuronans. Biochem Soc Trans. 1975;3(5):662–664. doi: 10.1042/bst0030662. [DOI] [PubMed] [Google Scholar]
  17. Sheehan J., Brass A., Almond A. The conformations of hyaluronan in aqueous solution: comparison of theory and experiment. Biochem Soc Trans. 1999 Feb;27(2):121–124. doi: 10.1042/bst0270121. [DOI] [PubMed] [Google Scholar]
  18. Staskus P. W., Johnson W. C., Jr Double-stranded structure for hyaluronic acid in ethanol-aqueous solution as revealed by circular dichroism of oligomers. Biochemistry. 1988 Mar 8;27(5):1528–1534. doi: 10.1021/bi00405a020. [DOI] [PubMed] [Google Scholar]
  19. Wik K. O., Comper W. D. Hyaluronate diffusion in semidilute solutions. Biopolymers. 1982 Mar;21(3):583–599. doi: 10.1002/bip.360210308. [DOI] [PubMed] [Google Scholar]

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