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Investigating the Secondary Structure of Membrane Peptides Utilizing Multiple 2H‑Labeled Hydrophobic Amino Acids via Electron Spin Echo Envelope Modulation (ESEEM) Spectroscopy
journal contribution
posted on 2018-04-03, 00:00 authored by Lishan Liu, Indra D. Sahu, Lauren Bottorf, Robert M. McCarrick, Gary A. LoriganAn
electron spin echo envelope modulation (ESEEM) approach was
used to probe local secondary structures of membrane proteins and
peptides. This ESEEM method detects dipolar couplings between 2H-labeled nuclei on the side chains of an amino acid (Leu
or Val) and a strategically placed nitroxide spin-label in the proximity
up to 8 Å. ESEEM spectra patterns for different samples correlate
directly to the periodic structural feature of different secondary
structures. Since this pattern can be affected by the side chain length
and flexibility of the 2H-labeled amino acid used in the
experiment, it is important to examine several different hydrophobic
amino acids (d3 Ala, d8 Val, d8 Phe)
utilizing this ESEEM approach. In this work, a series of ESEEM data
were collected on the AChR M2δ membrane peptide to build a reference
for the future application of this approach for various biological
systems. The results indicate that, despite the relative intensity
and signal-to-noise level, all amino acids share a similar ESEEM modulation
pattern for α-helical structures. Thus, all commercially available 2H-labeled hydrophobic amino acids can be utilized as probes
for the further application of this ESEEM approach. Also, the ESEEM
signal intensities increase as the side chain length gets longer or
less rigid. In addition, longer side chain amino acids had a larger 2H ESEEM FT peak centered at the 2H Larmor frequency
for the i ± 4 sample when compared to the corresponding i ± 3 sample. For shorter side chain amino acids, the 2H ESEEM FT peak intensity ratio between i ± 4 and i ± 3 was not well-defined.
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side chainAChR M 2δ membrane peptide2 H ESEEM FT peakacidESEEM signal intensities increaseElectron Spin Echo Envelope Modulation2 H-labeledside chain lengthESEEM modulation patternα- helical structures2 H Larmor frequency2 H ESEEM FT peak intensity ratioecho envelope modulation2 H-labeled nucleiESEEM approach8 Å. ESEEM spectra patterns
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