Protein Conformational Studies

A variety of bioanalytical techniques such as circular dichroism, analytical ultracentrifugation or gel filtration can be used to assess the conformational states of soluble proteins. Hydrodynamic measurements of reasonably pure samples by gel filtration HPLC or analytical ultracentrifugation can provide important information regarding changes in a protein's size, shape or interaction with ligands or other proteins. SPR biosensing can be used to measure kinetic interaction rates between proteins or ligands and the effects of mutants on protein conformation evaluated. The isolation of protein complexes from cells/tissues by co-immunoprecipitation followed by tryptic digestion and mass spectrometry provides a useful technique for identifying and characterizing stable interactions. Generating chemically-tagged protein constructs for /in vitro/ studies (eg., FRET, TRF etc) on protein-protein interactions or measurements of catalytically-induced changes in protein conformation is another aspect of the work Larial performs on behalf of its clients. Membrane proteins are often more difficult to study with respect to changes in their conformation. Methods such as partial proteolytic digestion followed by purification and peptide mapping of cleavage sites by mass spectrometry can be used to map topological changes in aqueous-exposed regions.



Protein molecular models enable predictive identification of aqueous-exposed sequences as potentially
suitable epitopes for synthetic antibody development programs