HFIP-induced structures and assemblies of the peptides from the transmembrane domain 4 of membrane protein Nramp1

Abstract

Membrane protein Nrampl (natural resistance-associated macrophage protein 1) is a pH-dependent divalent metal cation transporter that regulates macrophage activation in infectious and autoimmune diseases. A naturally occurring glycine to aspartic acid substitution at position 169 (G169D) within the transmembrane domain 4 (TM4) of Nrampl makes mice susceptible to Leishmania donovani, Salmonella typhimurium, and Mycobacterium bovis. Here we present a structural and self-assembling study on two synthetic 24-residue peptides, corresponding to TM4 of mouse Nrampl and its G169D mutant, respectively, in 1,1,1,3,3,3- hexafluoroisopropanol-d 2 (HFIP-d 2) aqueous solution by nuclear magnetic resonance (NMR) spectroscopy. The results show that amphipathic α-helical structures are formed from residue Ile173 to Tyr187 for the wild-type peptide and from Trp168 to Tyr187 for the G169D mutant, respectively. The segment of the N-terminus from Leu167 to Leu172 is poorly structured for the wild-type peptide, whereas it is well defined for the G169D mutant. Both peptides aggregate to form a tetramer and the monomeric peptides in peptide bundles are structurally and orientationally similar. The intermolecular interactions in assemblies could be stronger in the C-terminal regions related to residues Phe180-Leu184 than those in the central helical segments for both peptides. The G169D mutation may change the size of the opening mi the termini of assembly. © 2006 Wiley Periodicals, Inc.