Importance of physical vs. chemical interactions in surface shear rheology.
The stability of adsorbed protein layers against deformation has in literature been attributed to the formation of a continuous gel-like network. This hypothesis is mostly based on measurements of the increase of the surface shear elasticity with time. For several proteins this increase has been attributed to the formation of intermolecular disulfide bridges between adsorbed proteins. However, according to an alternative model the shear elasticity results from the low mobility of the densely packed proteins. To contribute to this discussion, the actual role of disulfide bridges in interfacial layers is studied. Ovalbumin was thiolated with S-acetylmercaptosuccinic anhydride (S-AMSA), followed by removal of the acetylblock on the sulphur atom, resulting in respectively blocked (SX) and deblocked (SH) ovalbumin variants. This allows comparison of proteins with identical amino acid sequence and similar globular packing and charge distribution, but different chemical reactivity. The presence and reactivity of the introduced, deblocked sulfhydryl groups were confirmed using the sulfhydryl-disulfide exchange index (SEI). Despite the reactivity of the introduced sulfhydryl groups measured in solution, no increase in the surface shear elasticity could be detected with increasing reactivity. This indicates that physical rather than chemical interactions determine the surface shear behaviour. Further experiments were performed in bulk solution to study the conditions needed to induce covalent aggregate formation. From these studies it was found that mere concentration of proteins (to 200 mg/mL, equivalent to a surface concentration of around 2 mg/m(2)) is not sufficient to induce significant aggregation to form a continuous network. In view of these results, it was concluded that the adsorbed layer should not be considered a gelled network of aggregated material (in analogy with three-dimensional gels formed from heating protein solutions). Rather, it would appear that the adsorbed proteins form a highly packed system of proteins with net-repulsive interactions.
Wierenga
Peter A
PA
Wageningen Centre for Food Sciences, P.O. Box 557, Diedenweg 20, 6700 AN, Wageningen, The Netherlands. peter.wierenga@wur.nl
Kosters
Hans
H
Egmond
Maarten R
MR
Voragen
Alphons G J
AG
de Jongh
Harmen H J
HH
D000327:Adsorption; D000818:Animals; D002645:Chickens; D004220:Disulfides; D004548:Elasticity; D010047:Ovalbumin / Q000737:chemistry*; D011487:Protein Conformation; D012212:Rheology; D013499:Surface Properties
D004220:Disulfides; D010047:Ovalbumin