title: Application of recent advances in hydrodynamic methods for characterising mucins in solution.
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 × 10(6) 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 × 10(6) 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 L p ~10 nm and mass per unit length, M L ~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.
authors.affiliation: National Centre for Macromolecular Hydrodynamics, University of Nottingham, Sutton Bonington, LE12 5RD, UK. stxfa7@nottingham.ac.uk.
authors.forename: Fahad M
authors.initials: FM
authors.lastname: Almutairi
authors.affiliation: Department of Physical Chemistry, University of Murcia, Regional Campus of Excellence Mare Nostrum, 30071, Murcia, Spain.
authors.forename: Jose-Gines Hernandez
authors.initials: JG
authors.lastname: Cifre
authors.affiliation: National Centre for Macromolecular Hydrodynamics, University of Nottingham, Sutton Bonington, LE12 5RD, UK.|School of Health Sciences, Faculty of Medicine and Health Science, University of Nottingham, Clifton Boulevard, Nottingham, NG7 2RD, UK.
authors.forename: Gary G
authors.initials: GG
authors.lastname: Adams
authors.affiliation: Department of Food Engineering, Abant Izzet Baysal University, Bolu, Turkey.
authors.forename: M Samil
authors.initials: MS
authors.lastname: Kök
authors.affiliation: Institute of Food Research, Norwich Research Park, Colney Lane, UK.
authors.forename: Alan R
authors.initials: AR
authors.lastname: Mackie
authors.affiliation: Department of Physical Chemistry, University of Murcia, Regional Campus of Excellence Mare Nostrum, 30071, Murcia, Spain.
authors.forename: Jose Garcia
authors.initials: JG
authors.lastname: de la Torre
authors.affiliation: National Centre for Macromolecular Hydrodynamics, University of Nottingham, Sutton Bonington, LE12 5RD, UK. steve.harding@nottingham.ac.uk.
authors.forename: Stephen E
authors.initials: SE
authors.lastname: Harding
mesh_terms: D000465:Algorithms; D000818:Animals; D031521:Fractionation, Field Flow / Q000379:methods; D057446:Hydrodynamics; D007413:Intestinal Mucosa / Q000378:metabolism; D055262:Mucin-2 / Q000737:chemistry*; D012996:Solutions; D013552:Swine
chemical_list: D055262:Mucin-2; D012996:Solutions
keywords: Conformation; Molar mass; Mucin; Novel hydrodynamic methods