ИСТИНА

An approach applied in this work involves the transmission electron microscopy of single macromolecules and molecular modeling. The Kv7.1 protein was expressed in P.pastoris, solubilized with detergent from cell membranes and purified on affinity column using the 1D4 epitope tag introduced on the C-terminus of polypeptide. Electron micrographs of protein samples were obtained with microscope JEOL 2100 and analyzed with EMAN 2 software. For the molecular modeling of the full-length structure of the Kv7.1 potassium channel we used its sequence similarity with the Kv1.2 channel (S1-S6 helices) which structure is known (PDB ID 3LUT). The Kv7.1 channel amino acid sequence analysis shows that its N- and C-terminal domains are disordered with the exception of C-terminal helical fragments of known structure (PDB IDs 4UMO, 3BJ4). Obtained model in general is in good accordance with 3D reconstruction based on electron microscopy data.