Self-Interacting Transmembrane Helices from the Human Single-Pass Membrane Proteome: The Impact of Primary Structure and Lipids on Affinity and Stoichiometry
Biophysics
Final Report Abstract
In sum, our results show that monomers, dimers, and trimers can be distinguished by measuring from steady-state anisotropies, provided that certain key parameters are determined by time-resolved measurements using a set of covalent reference proteins. The design of these reference proteins needs to correspond to the design of the candidate proteins. Varying the linker length between GFP moiety and oligomerization domain, for example, appears to strongly influence steady state anisotropy as shown here by comparing our sfGFP-based and EGF-based test cases. While soluble coiled-coil proteins have been used here to provide proof of principle, it is evident that the method can now be extended to integral membrane proteins. We expect that the application to membrane proteins proofs to be particularly valuable since the formers can be investigated in the membrane-reconstituted state (where other methods like size exclusion chromatography, native PAGE and analytical ultracentrifugation are not applicable). Unfortunately, solving the manifold technical and theoretical issues described in this report prevented us from investigating this further. These applications have to be reserved for future investigations.
Publications
- (2019) Experimental determination and data-driven prediction of homotypic transmembrane domain interfaces, Computational and Structural Biotechnology Journal 18 3230-3242
Xiao, Yao; Zeng, Bo; Berner, Nicola; Frishman, Dmitrij; Langosch, Dieter; Teese, Mark George
(See online at https://doi.org/10.1016/j.csbj.2020.09.035)
