title: In situ structural characterization of a recombinant protein in native Escherichia coli membranes with solid-state magic-angle-spinning NMR.
abstract: The feasibility of using solid-state magic-angle-spinning NMR spectroscopy for in situ structural characterization of the LR11 (sorLA) transmembrane domain (TM) in native Escherichia coli membranes is presented. LR11 interacts with the human amyloid precursor protein (APP), a central player in the pathology of Alzheimer's disease. The background signals from E. coli lipids and membrane proteins had only minor effects on the LR11 TM resonances. Approximately 50% of the LR11 TM residues were assigned by using (13)C PARIS data. These assignments allowed comparisons of the secondary structure of the LR11 TM in native membrane environments and commonly used membrane mimics (e.g., micelles). In situ spectroscopy bypasses several obstacles in the preparation of membrane proteins for structural analysis and offers the opportunity to investigate how membrane heterogeneity, bilayer asymmetry, chemical gradients, and macromolecular crowding affect the protein structure.
authors.affiliation: National High Magnetic Field Laboratory, Tallahassee, Florida 32310, United States.
authors.forename: Riqiang
authors.initials: R
authors.lastname: Fu
authors.forename: Xingsheng
authors.initials: X
authors.lastname: Wang
authors.forename: Conggang
authors.initials: C
authors.lastname: Li
authors.forename: Adriana N
authors.initials: AN
authors.lastname: Santiago-Miranda
authors.forename: Gary J
authors.initials: GJ
authors.lastname: Pielak
authors.forename: Fang
authors.initials: F
authors.lastname: Tian
mesh_terms: D004926:Escherichia coli / Q000166:cytology* / Q000235:genetics; D006801:Humans; D026502:LDL-Receptor Related Proteins / Q000032:analysis* / Q000235:genetics; D026901:Membrane Transport Proteins / Q000032:analysis* / Q000235:genetics; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D011487:Protein Conformation; D011993:Recombinant Fusion Proteins / Q000032:analysis* / Q000235:genetics
chemical_list: D026502:LDL-Receptor Related Proteins; D026901:Membrane Transport Proteins; D011993:Recombinant Fusion Proteins; C102582:SORL1 protein, human