A New Method for Determining the Local Environment and Orientation of Individual Side Chains of Membrane-Binding Peptides

Journal of the American Chemical Society

Volumn 126, Issue 16, 2004, Pages 5078-5079

  Tucker, Matthew J ^a   Getahun, Zelleka ^a   Nanda, Vikas ^a   DeGrado, William F ^a   Gai, Feng ^a  

^a UNIVERSITY OF PENNSYLVANIA   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

AMINO ACID; FLUORESCENT DYE; PEPTIDE;

AMINO ACID SEQUENCE; ANALYTIC METHOD; ARTICLE; CHEMICAL BOND; HYDRATION; MEMBRANE BINDING; MOLECULAR INTERACTION; NUCLEAR MAGNETIC RESONANCE SPECTROSCOPY; POLARIZATION; PROTEIN ANALYSIS; PROTEIN BINDING; PROTEIN STRUCTURE;

AMIDES; CELL MEMBRANE; HYDROPHOBICITY; LIPID BILAYERS; MAGNETIC RESONANCE SPECTROSCOPY; MEMBRANE PROTEINS; PEPTIDES; PHOSPHOLIPIDS; PROTEIN BINDING; PROTEIN STRUCTURE, SECONDARY; SPECTROPHOTOMETRY, INFRARED;

EID: 1942425119     PISSN: 00027863     EISSN: None     Source Type: Journal    
DOI: 10.1021/ja032015d     Document Type: Article

References (12)

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8. MPx-CNy peptides were synthesized by employing the standard Fmoc protocol, purified by reverse-phase HPLC, and verified by electrospray-ionization mass spectroscopy. For ATR-IR measurements, the MPx-CNy peptide (0.1 mM in MeOH) was added to POPC vesicles (15 mM in chloroform) in a ratio of 1:75 peptide/lipid. From this sample, 100 μL (∼650 μg) was then placed on a Ge crystal (5 x 1 cm) and allowed to dry. Before the experiment, the peptide-lipid sample was rehydrated by placing the Ge crystal in a humidified chamber for 12 h (covered beaker with water at the bottom and the Ge crystal supported to avoid immersion in water). Alternatively, rehydration was accomplished by placing 20 μL of water dropwise directly on the peptide/lipid bilayer surface. Both methods yielded similar results. ATR-IR spectra were collected with a Harrick's Horizon multiple-reflection attachment in conjunction with a Magna-IR 860 spectrometer equipped with a MCT detector. An average of 128 scans were taken for each sample.
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