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. The interactions between the attached glycans and the protein were assessed using the online LPC server (24)
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. Our efforts based on use of the EV limit mirror the use of the hardsphere fluid as a reference state for van der Waals liquids (51,52)
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. 2 should not be considered a binding curve, it may still be instructive to estimate an apparent average affinity of ANS for the four azurin species
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. Although for BSA a high binding capacity may be expected from its intrinsic tendency to bind nonpolar and aromatic moieties also in the liquid state, the above ANS binding data do suggest that beyond specific trends of individual polypeptides, multi-subunit proteins are more susceptible than monomers to destabilization and partial unfolding in the frozen state
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. An alternative explanation is that in this concentration range, the presence of an interfacial layer of protein has a marked influence on the observed rheology
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. We are grateful to Per Persson for initial assistance with electron microscopy work and to Jeffery Kelly and David Wemmer for providing the pmmHα plasmid
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. The stabilizing interactions themselves may be either native or nonnative, in the latter case suggesting cases of tailored nonnative interactions
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