Probing the dynamics of intact cells and nuclear envelope precursor membrane vesicles by deuterium solid state NMR spectroscopy.
Membrane dynamics is an essential part of many cellular mechanisms such as intracellular trafficking, membrane fusion/fission and mitotic organelle reconstitution. The dynamics of membranes is dependent primarily on their phospholipid and cholesterol composition and how these molecules are ordered in relation to one another. To determine the physical status of membranes in whole cells or purified membranes of subcellular compartments we have developed a novel application exploiting solid-state (2)H-NMR spectroscopy. We utilise this method to probe the dynamics of intact sperm and nuclear envelope precursor membranes. We show, using mass spectrometry, that either multilamellar or small unilamellar vesicles of deuterium-labelled palmitoyl-oleoylphosphatidylcholine can be used to probe the dynamics of sperm cells or nuclear envelope precursor membrane vesicles, respectively. Using (2)H-NMR we determine the order parameters of sperm cells and nuclear envelope precursor membrane vesicles. We demonstrate that whole sperm membranes are more dynamic than nuclear envelope precursor membranes due to the higher cholesterol levels of the latter. Our new application can be exploited as a generic method for monitoring membrane dynamics in whole cells, various subcellular membrane compartments and membrane domains in subcellular compartments.
Garnier-Lhomme
Marie
M
Cell Biophysics Laboratory, Lincoln's Inn Fields Laboratories, Cancer Research UK , 44, Lincoln's Inn Fields, WC2A 3PX London.
Grélard
Axelle
A
Byrne
Richard D
RD
Loudet
Cécile
C
Dufourc
Erick J
EJ
Larijani
Banafshé
B
D000731:Androstanes / Q000737:chemistry; D000818:Animals; D002462:Cell Membrane / Q000737:chemistry*; D002784:Cholesterol / Q000032:analysis; D002851:Chromatography, High Pressure Liquid; D003124:Colorimetry; D003903:Deuterium; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008297:Male; D009685:Nuclear Envelope / Q000737:chemistry*; D012617:Sea Urchins; D021241:Spectrometry, Mass, Electrospray Ionization; D013094:Spermatozoa
C415601:3-oxo-androstan-17-yl-(2'',2'',6'',6''-tetramethyl-N-oxyl)piperidyl butan-1',4'-dioate; D000731:Androstanes; D002784:Cholesterol; D003903:Deuterium