title: Application of REDOR subtraction for filtered MAS observation of labeled backbone carbons of membrane-bound fusion peptides.
abstract: Clean MAS observation of 13C-labeled carbons in membrane-bound HIV-1 and influenza fusion peptides was made by using a rotational-echo double-resonance spectroscopy (REDOR) filter of directly bonded 13C-15N pairs. The clean filtering achieved with the REDOR approach is superior to filtering done with sample difference spectroscopy. In one labeling approach, the peptide had labels at a single 13C carbonyl and its directly bonded 15N. The resulting chemical shift distribution of the filtered signal is used to assess the distribution of local secondary structures at the labeled carbonyl. For the influenza peptide, the Leu-2 carbonyl chemical shift distribution is shown to vary markedly with lipid and detergent composition, as well as peptide:lipid ratio, suggesting that the local peptide structure also has a strong dependence on these factors. Because most carboxylic- and amino-labeled amino acids are commercially available, this REDOR approach should have broad applicability to chemically synthesized peptides as well as bacterially synthesized proteins. In a second labeling approach, the HIV-1 fusion peptide had U-13C, 15N labeling over three sequential residues. When a 1.6 ms REDOR dephasing time is used, only backbone 13C signals are observed. The resulting spectra are used to determine spectral linewidths and to assess feasibility of assignment of uniformly labeled peptide.
authors.affiliation: Department of Chemistry, Michigan State University, East Lansing, MI 48824, USA.
authors.forename: Jun
authors.initials: J
authors.lastname: Yang
authors.forename: Paul D
authors.initials: PD
authors.lastname: Parkanzky
authors.forename: Michele L
authors.initials: ML
authors.lastname: Bodner
authors.forename: Craig A
authors.initials: CA
authors.lastname: Duskin
authors.forename: David P
authors.initials: DP
authors.lastname: Weliky
mesh_terms: D002247:Carbon Isotopes; D015497:HIV-1; D008561:Membrane Fusion; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D009975:Orthomyxoviridae; D017433:Protein Structure, Secondary; D014760:Viral Fusion Proteins / Q000737:chemistry*
chemical_list: D002247:Carbon Isotopes; D014760:Viral Fusion Proteins