title: T178 deletion impairs intermolecular interaction of the peptide Nramp1(164-191).
abstract: Natural resistance associated macrophage protein 1 (Nramp1), an integral membrane protein with 12 predicted transmembrane domains (TMs), is a divalent cation transporter associated with infectious and autoimmune diseases. A naturally occurring mutation G169D within TM4 of Nramp1 leads to the loss of function, suggesting potential importance of TM4 for the biological function of the protein. In this study, we determine the three-dimensional structure and topology of a synthetic peptide, del(T178), corresponding to Nramp1(164-191) (basically consisting of the putative TM4 of Nramp1) with Thr178 deletion in TFE and SDS micelles using NMR and CD spectroscopic techniques, and compare the results with those of the wildtype peptide. Similarly to the wildtype peptide, the del(T178) peptide still forms an amphiphilic-like alpha-helical structure in both membrane mimics and is embedded in SDS micelles. Differently, whereas the wild-type peptide forms a helix bundle with the hydrophilic side facing the interior of the bundle, the del(T178) peptide exists as a monomer in the membrane mimics and the hydrophilic side of the helix is located near the interface of SDS micelles. Moreover, a strongly cooperative protonation occurs between intramolecular Asp residues for the del(T178) peptide in SDS micelles, while the cooperative proton binding between intermolecular Asp residues was observed for the wildtype peptide. The difference in the results of the two peptides suggests that the deletion of Thr178 impairs intermolecular interaction of the peptide.
authors.affiliation: State Key Laboratory of Supramolecular Structure and Materials, Jilin University, Changchun 130012, People's Republic of China.
authors.forename: Rong
authors.initials: R
authors.lastname: Xue
authors.forename: Shuo
authors.initials: S
authors.lastname: Wang
authors.forename: Haiyan
authors.initials: H
authors.lastname: Qi
authors.forename: Yuande
authors.initials: Y
authors.lastname: Song
authors.forename: Shuyan
authors.initials: S
authors.lastname: Xiao
authors.forename: Fei
authors.initials: F
authors.lastname: Li
mesh_terms: D027682:Cation Transport Proteins / Q000737:chemistry* / Q000235:genetics; D002942:Circular Dichroism; D009682:Magnetic Resonance Spectroscopy; D008823:Micelles; D008958:Models, Molecular; D010455:Peptides / Q000737:chemistry*; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D017384:Sequence Deletion; D040681:Structural Homology, Protein; D000099173:Natural Resistance-associated Macrophage Protein 1
chemical_list: D027682:Cation Transport Proteins; D008823:Micelles; D010455:Peptides; D000099173:Natural Resistance-associated Macrophage Protein 1