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Related Experiment Videos

Using O2 to probe membrane immersion depth by 19F NMR.

R S Prosser1, P A Luchette, P W Westerman

  • 1Department of Chemistry, Kent State University, Kent, OH 44242; and Northeastern Ohio Universities' College of Medicine, P. O. Box 95, Rootstown, OH 44272, USA. sprosser@membrane.kent.edu

Proceedings of the National Academy of Sciences of the United States of America
|August 24, 2000
PubMed
Summary

This study shows that high-pressure oxygen can reveal the depth of fluorinated molecules within membranes using fluorine-19 NMR (19F NMR). This technique helps determine solvent accessibility and immersion depth of fluorinated groups in membrane-associated macromolecules.

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Area of Science:

  • Membrane biophysics
  • Nuclear Magnetic Resonance (NMR) spectroscopy
  • Fluorine chemistry

Background:

  • Lipid bilayers are crucial models for cell membranes.
  • Determining the precise location of molecules within membranes is essential for understanding their function.
  • Fluorine-19 NMR (19F NMR) offers a sensitive probe for fluorinated molecules.

Purpose of the Study:

  • To demonstrate the feasibility of using (19)F NMR to determine solvent accessibility and membrane immersion depth of fluorinated groups.
  • To investigate the effect of apolar oxygen on (19)F NMR relaxation rates.
  • To establish a method for probing the location of fluorinated species in membrane systems.

Main Methods:

  • Reconstitution of a fluorinated detergent (CF(3)(CF(2))(5)C(2)H(4)-O-maltose) into a lipid bilayer model membrane.

Related Experiment Videos

  • Application of (19)F NMR spectroscopy to analyze the fluorinated detergent.
  • Systematic variation of oxygen partial pressure (P(O(2))) up to 20 bar.
  • Measurement of spin-lattice relaxation rates (R(1)) and chemical shifts.
  • Comparison with results obtained using a membrane surface-associated paramagnetic species (CAT-16).

    • Main Results:

    • Apolar oxygen exerts paramagnetic relaxation effects on (19)F nuclei, dependent on immersion depth.
    • Increasing oxygen pressure to >20 bar significantly amplifies these effects with minimal line broadening.
    • Differences in longitudinal relaxation rates at high oxygen pressure correlate with the expected burial depth of fluorine groups.
    • This oxygen-induced effect contrasts with the trend observed with surface-associated paramagnetic agents.

    Conclusions:

    • High-pressure oxygen can be used as an effective tool to probe the membrane immersion depth and solvent accessibility of fluorinated groups in membrane-associated molecules via (19)F NMR.
    • Differential relaxation rates induced by oxygen provide a sensitive measure of molecular location within lipid bilayers.
    • This (19)F NMR-based approach offers a valuable method for studying membrane-associated macromolecules.