Protein conformation and interaction

New Insights into Protein Structure and Function

Direct structural measurement without tags

Valuable new insights into protein function by direct, real time measurements. Conformational changes associated with ions, pH, other proteins, or molecules are observed at a resolution of less than a tenth of a carbon-carbon bond length. Simultaneous measurements of mass, concentration and density provide a highly correlated perspective of activity, without radioactive or fluorescent tags.

The AnaLight® Bio200 is a new instrument with the sensitivity to directly measure proteins changing conformation, as they function. If the function includes mass capture (such as ligand or small molecule binding) there is usually an increase in density, as the binding partner disturbs the site solvation environment. The system has the exquisite sensitivity required to detect such density changes. This capability to simultaneously detect binding of small molecules, whilst measuring conformational changes, provides a truly unique and highly correlated insight into protein structure, function and mechanism.

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Key applications

The Farfield Biomolecular Characterisation Matrix demonstrates the wealth of data that is available by simultaneously quantifying thickness and density at molecular resolutions. Conventional methods rely solely on a mass change, whereas the thickness/density matrix highlights 9 unique fingerprints. These can be associated to specific protein functions as both thickness and density can be either decreasing, () constant, (-) or increasing (). Quite often the stimulus is largely mass neutral, as with small molecule studies or protein folding due to ion or pH changes. The only way to detect such functions without recourse to labels is to measure one or ideally both of these parameters

An example of the exquisite resolution of the AnaLight® Bio200 is where an enzyme (transglutaminase) conformational change of 0.1nm-0.4nm is quantified in real time at different concentrations of Ca2+ ion with a resolution of better than 0.01nm.

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