Integrated Structural Biology for α-Helical Membrane Protein Structure Determination

摘要: While great progress has been made, only 10% of the nearly 1,000 integral, α-helical, multi-span membrane protein families are represented by at least one experimentally determined structure in PDB. Previously, we developed algorithm BCL::MP-Fold, which samples large conformational space proteins de novo assembling predicted secondary elements guided knowledge-based potentials. Here, present a case study rhodopsin fold determination integrating sparse and/or low-resolution restraints from multiple experimental techniques including electron microscopy, paramagnetic resonance spectroscopy, and nuclear magnetic spectroscopy. Simultaneous incorporation orthogonal not significantly improved sampling accuracy but also allowed identification correct fold, is demonstrated size-normalized transmembrane root-mean-square deviation as low 1.2 A. The protocol this can be used for unknown folds when limited available.