Åke Wieslander

Water binding and phase structures for different Acholeplasma laidlawii membrane lipids studied by deuteron nuclear magnetic resonance and x-ray diffraction

Water binding capability and phase structures for different lipid species extracted from Acholeplasma laidlawii A membranes have been studied using deuteron nuclear magnetic resonance and low-angle X-ray diffraction. The dominating membrane lipids are monoglucosyldiglyceride and diglucosyldiglyceride and each of them takes up limited amounts of water (bound plus trapped), i.e., up to 13% (w/w), whereas the phospholipids and phosphoglycolipids have larger hydration capacities. Addition of magnesium and calcium ions, but not sodium ions, to the diglucosyldiglyceride increases the hydration capability. This increase is accompanied by the formation of a metastable liquid crystalline phase and a hysteresis effect for the transition temperature. Large differences in water deuteron quadrupole splitting were observed between mono- and diglucosyldiglyceride. Both 2H nuclear magnetic resonance and low-angle X-ray diffraction studies on lipids containing biosynthetically incorporated omega-d3-palmitic acid clearly indicate the existence of a reverse hexagonal phase structure for the monoglucosyldiglyceride and lamellar structures for the diglucosyldiglyceride and the other membrane lipids. The low hydration capability of the large diglucosyldiglyceride polar head is discussed in terms of polar head configuration. Both mono- and diglucosyldiglyceride have several physical properties similar to those of phosphatidylethanolamine.

Qualitative and quantitative variations of membrane lipid species in Acholeplasma laidlawii A

In Acholeplasma laidlawii A, strain EF 22, the relative amounts of the membrane polar lipids vary as a consequence of different fatty acid supplements to the growth medium. The number of lipid species also varies; a new apolar monoglucolipid containing four fatty acid residues was present only when saturated fatty acids dominated in the growth medium. A new phosphoglucolipid, probably with a glycerophosphoryl-monoglucosyldiglyceride structure, was also found. The most pronounced variations occurred between the two dominating glucolipids, monoglucosyldiglyceride and diglucosyldiglyceride; the former being found in larger amounts when a saturated or a trans-unsaturated fatty acid was present in the medium. The amount of diglucosyldiglyceride decreased accordingly. A qualitative relationship between fatty acid properties and membrane lipid variations was established over a wide fatty acid concentration range. Incorporation of supplied fatty acids reached higher levels than normally found in other acholeplasmas. The ratio between membrane protein and lipids exhibited significant and coherent variations during growth and was to some extent influenced by the fatty acids in the medium. These changes indicate variations in lipid-protein organization in the membranes during growth.

Lipid phase structure governs the regulation of lipid composition in membranes of acholeplasma laidlawii

great variety of lipid molecules are present in biologic membr~es - a rn~rn~an cell membrane may contain 100 different lipids including intra- molecular variants. Many of these form lamellar liquid crystalline phases together with water [ I]_ However, most biological membr~es contain at least one major lipid species forming a non-lamellar phase. Monog~actosyl~~y~~de and mono~ucosyldi- glyceride (MGDG) form a reversed hexagonal (HE) phase [ 2,3], while

Control of membrane polar lipid composition in Acholeplasma laidlawii a by the extent of saturated fatty acid synthesis

The low level of endogenous fatty acid synthesis in Acholeplasma laidlawii A strain EF22 was found to be caused by a deficiency of pantetheine in the lipid-depleted growth medium. By supplementing the oleic acid-containing medium with increasing concentrations of pantethein, saturated fatty acid synthesis was stimulated (having an apparent Km of 5 microM for pantetheine) and the incorporation of endogenously synthesized fatty acids in membrane lipids increased markedly. Furthermore, carotenoid biosynthesis was stimulated. Exogenous palmitic acid was found to inhibit partially the endogenous fatty acid synthesis. A gradual stimulation of fatty acid synthesis was accompanied by a linear increase in the molar proportion between the two dominating membrane glucolipids, monoglucosyldiacylglycerol and diglucosyldiacylglycerol. The total amount of charged membrane lipids decreased upon increasing the degree of fatty acid saturation. These regulations are discussed in terms of membrane stability, and influence of membrane molecular ordering and surface charge density on lipid polar head group synthesis.

Orientation of β-carotene and retinal in lipid bilayers

The function of carotenoids in membranes of photosynthetic bacteria and plants has been intensively investigated [l-4]. In animals certain carotenoids such as p-carotene fullfil an essential physiological function as precursors of vitamin A. By enzymatic oxidation p-carotene is converted to retinol which is further oxidized to retinal i.e., vitamin A. The positioning of carotenoids in membranes is far from clear [ .5,6]. No conclusive experiment has been performed showing the orientation of any carotenoid in lipid bilayers or biological membranes. Here, we have used polarized light spectroscopy and studied the orientation of p-carotene and retinal in different types of lipid bilayers.

Fractionation of membranes from Acholeplasma laidlawii A on the basis of their surface properties by partition in two-polymer aqueous phase systems

Acholeplasma laidlawii A consists of pleomorphic cell clusters surrounded by a single membrane. When lysed, a cell gives rise to several membrane fragments which cannot be separated from each other by isopycnic sucrose gradient centrifugation. A heterogeneous lateral organization of the cell membranes was detected by countercurrent distribution of membrane fragments in a two-polymer aqueous phase system. It revealed that the membranes consist of at least two subpopulations with respect to surface properties. Changes in the fatty acid and cholesterol content of the membranes revealed that the resolution of different subpopulations was predominantly due to a critical ratio of monoglucosyldiglyceride to diglucosyldiglyceride. The heterogeneity of the membrane probably depends on lipid-lipid and lipid-protein steric interactions. Charged lipids, an apolar monoglucolipid and the ratio between lipids and proteins also affect membrane partition. The differences in the subpopulations were further reflected by different specific activities of NADH dehydrogenase, NADH oxidase and ATPase. These activities varied independently. Minor quantitative differences in the protein patterns of different subpopulations were apparent. The origin and the preservation of the membrane subpopulations are discussed in terms of lipid-lipid and lipid-protein interactions, their age and energy metabolism.

Effects of anesthetics on water permeability and lipid metabolism in acholeplasma laidlawii membranes

The addition of tetracaine and diethyl ether to Acholeplasma laidlawii at concentrations commonly used for local anesthesia did not affect water permeability over the cell membrane, as measured by a pulsed magnetic field gradient NMR method. However, A. laidlawii changed its membrane lipid composition upon treatment with these anesthetics. Both tetracaine and diethyl ether addition resulted in a decrease in the molar ratio between the major membrane glucolipids, monoglucosyldiacylglycerol and diglucosyldiacylglycerol. The ratio between saturated and unsaturated acyl chains did not change. The results are in accordance with our proposal that A. laidlawii regulates its lipid composition in order to maintain optimal packing stability in the membrane (Wieslander, A., Christiansson, A., Rilfors. L. and Lindblom, G. (1980) Biochemistry 19, 3650--3655). Introduction of anesthetics into the hydrophobic region of a bilayer is likely to affect the lipid packing. A membrane which contains lipids like monoglucosyldiacylglycerol, which forms a reversed hexagonal phase, will be destabilized unless the amounts of such lipids are reduced. The membrane concentration of anesthetics was estimated to one molecule per 12--15 lipid molecules. The fact that A. laidlawii regulates its lipid composition as a response to these concentrations, despite their negligible effect on water permeability, indicates a high sensitivity of this regulatory system.