Application of recent advances in hydrodynamic methods for characterising mucins in solution

Almutairi, Fahad M. and Cifre, Jose-Gines Hernandez and Adams, Gary G. and Kök, M. Samil and Mackie, Alan R. and de la Torre, Jose Garcia and Harding, Stephen E. (2016) Application of recent advances in hydrodynamic methods for characterising mucins in solution. European Biophysics Journal, 45 (1). pp. 45-54. ISSN 1432-1017

[img] PDF - Repository staff only - Requires a PDF viewer such as GSview, Xpdf or Adobe Acrobat Reader
Download (5MB)

Abstract

Mucins are the primary macromolecular component of mucus—nature’s natural lubricant—although they are poorly characterised heterogeneous substances. Recent advances in hydrodynamic methodology now offer the opportunity for gaining a better understanding of their solution properties. In this study a combination of such methods was used to provide increased understanding of a preparation of porcine intestinal mucin (PIM), MUC2 mucin, in terms of both heterogeneity and quantification of conformational flexibility. The new sedimentation equilibrium algorithm SEDFIT-MSTAR is applied to yield a weight average (over the whole distribution) molar mass of 7.1 × 106 g mol−1, in complete agreement with size exclusion chromatography coupled with multi-angle light scattering (SEC-MALS), which yielded a value of 7.2 × 106 g mol−1. Sedimentation velocity profiles show mucin to be very polydisperse, with a broad molar mass distribution obtained using the Extended Fujita algorithm, consistent with the elution profiles from SEC-MALS. On-line differential pressure viscometry coupled to the SEC-MALS was used to obtain the intrinsic viscosity [η] as a function of molar mass. These data combined with sedimentation coefficient data into the global conformation algorithm HYDFIT show that PIM has a flexible linear structure, with persistence length Lp ~10 nm and mass per unit length, ML ~2380 g mol−1 nm−1, consistent with a Wales-van Holde ratio of ~1.2 obtained from the concentration dependence of the sedimentation coefficient.

Item Type: Article
Additional Information: The final publication is available at Springer via http://dx.doi.org/10.1007/s00249-015-1075-0
Keywords: Mucin; Molar mass; Conformation; Novel hydrodynamic methods
Schools/Departments: University of Nottingham, UK > Faculty of Science > School of Biosciences
Identification Number: 10.1007/s00249-015-1075-0
Depositing User: Eprints, Support
Date Deposited: 10 May 2017 13:34
Last Modified: 10 May 2017 21:44
URI: http://eprints.nottingham.ac.uk/id/eprint/42720

Actions (Archive Staff Only)

Edit View Edit View