High-resolution proton NMR measures mobile lipids associated with Triton-resistant membrane domains in haematopoietic K562 cells lacking or expressing caveolin-1.
High-resolution proton NMR spectra of intact tumour cells generally exhibit intense signals due to isotropically mobile lipids (MLs) of still uncertain nature and origin. NMR studies performed on intact wild-type and caveolin-1-infected haematopoietic K562 cells showed that, under our experimental conditions, part of the ML signals are due to lipid complexes resistant to extraction in Triton X-100 at 4 degrees C. This evidence suggests that a portion of NMR-visible lipid structures are compatible with Triton-resistant membrane rafts and therefore biophysically distinct from NMR-visible Triton-soluble lipid bodies. Similarly to lipid rafts and caveolae, the organization of the Triton-insoluble ML domains could be compromised by treatment with beta-octylglucoside or methyl-beta-cyclodextrin. Exposure to exogenous sphingomyelinase caused an increase in ML NMR visibility, indicating the possible involvement of ceramides in ML formation. The mobility of these lipids was found to be temperature sensitive, suggesting a transition in cells going from 4 degrees C to 25-37 degrees C. These new results are here discussed in the light of possible contributions of plasma membrane microdomains to NMR-visible ML signals.
Ferretti
A
A
Laboratory of Cell Biology, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, Italy.
Knijn
A
A
Raggi
C
C
Sargiacomo
M
M
D051242:Caveolin 1; D022461:Caveolins / Q000737:chemistry / Q000172:deficiency / Q000378:metabolism*; D003505:Cyclodextrins / Q000494:pharmacology; D004351:Drug Resistance; D005960:Glucosides / Q000494:pharmacology; D006412:Hematopoietic Stem Cells / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D020014:K562 Cells; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008560:Membrane Fluidity / Q000187:drug effects; D008563:Membrane Lipids / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism*; D021962:Membrane Microdomains / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D017830:Octoxynol / Q000737:chemistry* / Q000494:pharmacology*; D011522:Protons; D012995:Solubility; D013108:Sphingomyelin Phosphodiesterase / Q000494:pharmacology; D013696:Temperature; D047392:beta-Cyclodextrins
C495540:CAV1 protein, human; D051242:Caveolin 1; D022461:Caveolins; D003505:Cyclodextrins; D005960:Glucosides; D008563:Membrane Lipids; D011522:Protons; D047392:beta-Cyclodextrins; C108732:methyl-beta-cyclodextrin; C018619:octyl-beta-D-glucoside; D017830:Octoxynol; D013108:Sphingomyelin Phosphodiesterase