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Updated: Jun 4, 2026

Mucin Agarose Gel Electrophoresis: Western Blotting for High-molecular-weight Glycoproteins
Published on: June 14, 2016
Normal and shear forces between surfaces bearing porcine gastric mucin, a high-molecular-weight glycoprotein
Neale M Harvey1, Gleb E Yakubov, Jason R Stokes
1The Physical and Theoretical Chemistry Laboratory, University of Oxford, South Parks Road, Oxford OX1 3QZ, United Kingdom.
Abstract:
A surface force balance was used to measure the normal and shear forces between two mica surfaces each bearing an adsorbed layer of porcine gastric mucin ("Orthana" mucin), genetically similar to human MUC6. This mucin is a highly purified, 546 kDa, weakly negative, polyampholytic molecule with a "dumbbell" structure. Both bare (HP) and hydrophobized (HB) mica substrates were used, and forces were measured under 1 and 30 mg/mL mucin solutions, under pure (no-added-salt) water, and under 0.1 M aqueous Na(+) solution. Normal surface forces were monotonically repulsive in all cases, with onset of repulsion occurring at smaller surface separations, D, in the 0.1 M salt solutions (∼ 20 nm, compared with ∼40 nm for no added salt). Repulsion on HP mica was greater on surface compression than decompression, an effect, attributed to bridging and slow-relaxing additional adsorption on compression, not seen on HB mica, a difference attributed to the denser coverage of mucin hydrophobic moieties on the HB surface. Friction forces increased with compression in all cases, showing hysteretic behavior on HP but not on HB mica, commensurate with the hysteresis observed in the normal measurements. Low friction coefficients μ (= ∂F(s)/∂F(n) < 0.05) were seen up to mean pressures
≈ 0.5 to 1.0 MPa, attributed to low interpenetration of the opposed layers together with hydration lubrication effects, with higher μ (up to 0.4) at higher
attributed to interlayer entanglements and to bridging (for the case of HP mica). Shear forces increased only weakly with sliding speed over the range investigated (80-820 nm s(-1)). The lower friction with HB relative to HP mica suggests a selectivity of the HB surface to the hydrophobic moieties of the mucin that in consequence exposes relatively more of the better-lubricating hydrophilic groups. This surface-selectivity effect on lubrication may have a generality extending to other biological macromolecules that contain both hydrophilic and hydrophobic groups.
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