PCS-based structure determination of protein–protein complexes

• DOI: 10.1007/s10858-010-9401-4
• Publication Date: 2010-03-18T03:30:29Z
• Authors: Tomohide Saio; Masashi Yokochi; Hiroyuki Kumeta; Fuyuhiko Inagaki
• Keywords: Heteronuclear molecule; Chemical shift; Docking (animal)

A simple and fast nuclear magnetic resonance method for docking proteins using pseudo-contact shift (PCS) 1HN/15N chemical perturbation is presented. PCS induced by a paramagnetic lanthanide ion that attached to target protein binding peptide tag anchored at two points. provides long-range (~40 Å) distance angular restraints between the observed nuclei, while data provide loose contact-surface information. The usefulness of this was demonstrated through structure determination p62 PB1-PB1 complex, which forms front-to-back 20 kDa homo-oligomer. As PB1 does not intrinsically bind metal ions, one subunit dimer anchoring Each monomer treated as rigid body docked based on backbone data. Unlike NOE-based structural determination, only requires assignments signals obtained from several sets two-dimensional 15N-heteronuclear single quantum coherence spectra, thus facilitating rapid protein–protein complex.