In-depth interrogation of protein thermal unfolding data with MoltenProt.

ABSTRACT: Protein stability is a key factor in successful structural and biochemical research. However, the approaches for systematic comparison of protein stability are limited by sample consumption or compatibility with sample buffer components. Here we describe how miniaturized measurement of intrinsic tryptophan fluorescence (NanoDSF assay) in combination with a simplified description of protein unfolding can be used to interrogate the stability of a protein sample. We demonstrate that improved protein stability measures, such as apparent Gibbs free energy of unfolding, rather than melting temperature T_m , should be used to rank the results of thermostability screens. The assay is compatible with protein samples of any composition, including protein complexes and membrane proteins. Ou

SUBMITTER: Kotov V 

PROVIDER: S-EPMC7737771 | biostudies-literature | 2021 Jan

REPOSITORIES: biostudies-literature