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Measuring the effect of ligand binding on the interface stability of multimeric proteins using dynamic light scattering.

ABSTRACT: We have demonstrated that an approach using guanidine hydrochloride at low concentrations to progressively disrupt protein-protein interactions can be quantitated using dynamic light scattering. This approach is sensitive enough to detect ligand-induced changes of subunit-subunit interactions for homo-hexameric glutamate dehydrogenase, allowing ΔΔG of reversible subunit dissociation to be calculated. The use of dynamic light scattering makes this approach generally applicable to soluble proteins to monitor the relative strength of protein-protein interactions with a particular emphasis on assessing the impact of ligand binding on such interfaces.

SUBMITTER: Marion JD

PROVIDER: S-EPMC2949477 | biostudies-other | 2010 Dec

REPOSITORIES: biostudies-other

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Measuring the effect of ligand binding on the interface stability of multimeric proteins using dynamic light scattering.

Marion James D JD Van Danielle N DN Bell J Ellis JE Bell Jessica K JK

Analytical biochemistry 20100821 2

We have demonstrated that an approach using guanidine hydrochloride at low concentrations to progressively disrupt protein-protein interactions can be quantitated using dynamic light scattering. This approach is sensitive enough to detect ligand-induced changes of subunit-subunit interactions for homo-hexameric glutamate dehydrogenase, allowing ΔΔG of reversible subunit dissociation to be calculated. The use of dynamic light scattering makes this approach generally applicable to soluble proteins ...[more]

PMID: 20732291

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