Abstract

Fluorescence polarization of cis- and trans-parinaric acid and high angle x-ray diffraction are used to examine the physical effects of in vitro sterol depletion of LM cell plasma membranes. Measurements on lipids isolated from the normal plasma membrane show that most of the lipid is fluid between 5 and 45 degrees C. When no sterol is present, approximately 15% of the phospholipids undergo gel phase formation, detectable by x-ray measurements, in this temperature range. Partial sterol repletion of the isolated phospholipids serves to lower the onset temperature of the phase transition, as well as to decrease the fraction of lipid undergoing a transition. Intact plasma membranes are sterol depleted by in vitro incubation with egg phosphatidylcholine liposomes. This induces a phase transition with an onset temperature increasingly above the physiological temperature as more sterol is removed from the membranes. The transition can be suppressed by returning sterol to the membranes. These experiments extend earlier work (Rintoul, D. A., Chou, S.-M., and Silbert, D. F. (1979) J. Biol. Chem. 254, 10070-10077) on isolated plasma membrane lipids by showing that sterol suppresses a phase transition in intact membranes as well as in isolated lipids. The correlation between gel phase formation upon sterol loss at the physiological temperature and loss of membrane glucose transport function is discussed.

Highlights

  • Fluorescence polarizationof cis- and trans-parinaric cases, less well defined than in the studies of Thilo et al

  • Acids and high angle x-ray diffraction are usedto ex- (1977)in which transport activities below and above both end amine the physical effects of in vitro sterol depletioonf points of the transition were determined to show that the LM cellplasmamembranes.Measurementsonlipids sugar transporterspartition preferentially into fluid phase isolated from the normal plasma membrane show thatlipid

  • Chem. 254,10070-10077) onisolated The data indicate that sterol depletion is associated with the plasma membrane lipids by showing that sterol sup- appearance of a gel to liquid crystalline transition, which presses a phase transition in intact membranaesswell begins above the physiological temperature, and involves a as in isolated lipids

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Summary

Full size photocopies are available from the Journal of Biological

‘The abbreviations used are: P, the polarization ratio, P = Al/Zl with the values corrected for instrument anisotropy; tPnA, 9, 11, 13, 15- all-trans-octadecatetraenoicacid; cPnA, 9, 11, 13, 15-cis, trans, ument No 81M-229, cite author(s), and include a check or money order for $2.80 per set of photocopies. Full size photocopies are also included in the microfilm edition of the Journal thaits available from Waverly Press.

The Physical Role of Sterionl a Mammalian Plasma Membrane
DISCUSSION
ThPehysical Role of Sterol inMaammaliaPnlasma
Findings
The PhysicalRole ofSterol in a Mammalian PlasmMaembrane
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