title: A Guide to Differential Scanning Calorimetry of Membrane and Soluble Proteins in Detergents.
abstract: Differential scanning calorimetry (DSC) detects protein thermal unfolding by directly measuring the heat absorbed. Simple DSC experiments that require relatively small amounts of pure material can provide a wealth of information related to structure, especially with respect to domain architecture, without the need for a complete thermodynamic analysis. Thus, DSC is an ideal additional tool for membrane protein characterization and also offers several advantages over indirect thermal unfolding methods. Integral membrane proteins (IMPs) that comprise both large multitopic transmembrane domains (TMDs) and extramembranous domains (EMDs) are differentially affected by detergent interactions with both domains. In fact, in some cases, destabilization of the EMD by detergent may dominate overall IMP stability. This chapter will (1) provide a perspective on the advantages of DSC for membrane protein characterization and stability measurements, including numerous examples spanning decades of research; (2) introduce models for the interaction and destabilization of IMPs by detergents; (3) discuss two case studies from the authors' lab; and (4) offer practical advice for performing DSC in the presence of detergents.
authors.affiliation: Center for Structural Biology, University of Alabama at Birmingham, Birmingham, Alabama, USA.
authors.forename: Zhengrong
authors.initials: Z
authors.lastname: Yang
authors.affiliation: Center for Structural Biology, University of Alabama at Birmingham, Birmingham, Alabama, USA; Department of Chemistry, University of Alabama at Birmingham, Birmingham, Alabama, USA. Electronic address: christie@uab.edu.
authors.forename: Christie G
authors.initials: CG
authors.lastname: Brouillette
mesh_terms: D002152:Calorimetry, Differential Scanning / Q000379:methods*; D003902:Detergents / Q000737:chemistry*; D008565:Membrane Proteins / Q000737:chemistry*; D058767:Protein Unfolding; D013816:Thermodynamics
chemical_list: D003902:Detergents; D008565:Membrane Proteins
keywords: CFTR; DSC; Detergent; Differential scanning calorimetry; Membrane protein; Membrane protein stability; NBD1; Thermal stability; Thermal unfolding