The atomic-level mechanism underlying the functionality of aquaporin-0

Abstract

So far, more than 82,000 protein structures have been reported in the Protein Data Bank, but the driving force and structures that allow for protein functions have not been elucidated at the atomic level for even one protein. We have been able to clarify that the inter-subunit hydrophobic interaction driving the electrostatic opening of the pore in aquaporin 0 (AQP0). Aquaporins are membrane channels for water and small non-ionic solutes found in animals, plants, and microbes. The structures of aquaporins have high homology and consist of homotetramers, each monomer of which has one pore for a water channel. Each pore has two narrow portions: one is the narrowest constriction region consisting of aromatic residues and an arginine (ar/R), and another is two asparagine-proline-alanine (NPA) homolog portions. Here we show that an inter-subunit hydrophobic interaction in AQP0 drives a stick portion consisting of four amino acids toward the pore and the tip of the stick portion, consisting of a nitrogen atom, opens the pore: that movement is the swing mechanism (this http URL). The energetics and conformational change of amino acids participating in the swing mechanism confirm this view. The swing mechanism in which inter-subunit hydrophobic interactions in the tetramer drive the on-off switching of the pore explains why aquaporins consist of tetramers. Here, we report that experimental and molecular dynamics findings using various mutants support this view of the swing mechanism. The finding that mutants of amino acids in AQP2 corresponding to the stick of the swing mechanism cause severe recessive nephrogenic diabetes insipidus (NDI) demonstrates the critical role of the swing mechanism for the aquaporin function. We report first that the inter-subunit hydrophobic interaction in aquaporin 0 drives the electrostatic opening of the aquaporin pore at the atomic level.