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J R Silvius

Publications and source records attributed to J R Silvius.

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Lipid compositional manipulation in Acholeplasma laidlawii B. Effect of exogenous fatty acids on fatty acid composition and cell growth when endogenous fatty acid production is inhibited.

A variety of potential inhibitors of de novo fatty acid biosynthesis have been tested for activity in Acholeplasma laidlawii B. Two compounds, avidin and N,N-dimethyl-4-oxo-2trans-dodecenamide (CM-55), an antimicrobial fatty amide, strongly inhibit de novo biosynthesis without nonspecific toxic effects at moderate dosages. Avidin is the more potent inhibitor, abolishing de novo fatty acid synthesis and greatly reducing the chain elongation of exogenous fatty acids at level of 25 U/l. CM-55 gives complete inhibition of de novo biosynthesis only at low temperatures and inhibits exogenous fatty acid elongation to a variable extent. However, CM-55 is still a more potent antilipogenic agent in this organism than is the fungal antibiotic cerulenin. Cells cultured with avidin grow only when one or more exogenous medium- or long-chain fatty acids are added to the growth medium. The extent of cell growth under these conditions depends primarily on the physical properties of the exogenous fatty acid(s). In general, fatty acids giving diacylglycerolipids of very high or very low fluidity are unsuitable growth substrates, while those whose diacylglycerol derivatives are of intermediate fluidity support fair to good cell growth.

Acholeplasma laidlawii↗

Membrane lipid biosynthesis in Acholeplasma laidlawii B: de novo biosynthesis of saturated fatty acids by growing cells.

The de novo biosynthesis of fatty acids of 12 to 18 carbons from precursors of 5 carbons or fewer has been demonstrated in Acholeplasma laidlawii B. Radiolabeling experiments indicated that the normal primers for the synthesis of the even- and odd-chain fatty acids are acetate and propionate or valerate, respectively. Saturated straight-chain monomethyl-branched fatty acids of up to five carbons were readily utilized as primers, wheras more highly branched species and those possessing halogen substituents or unsaturation were not utilized. At primer concentrations of 1 to 3 mM, up to 80% of the total cellular lipid fatty acids were derived from exogenous primer. The mean chain length of the exogenous primer-derived fatty acids rose with increasing primer incorporation for methyl-branched short-chain fatty acids but was invariant for propionate. The products of de novo biosynthesis varied only slightly with temperature or cholesterol supplementation, suggesting that de novo biosynthesis is not directly influenced by membrane fluidity. Cerulenin inhibited de novo biosynthesis in a fashion that suggests the presence of two beta-ketoacyl thioester synthetases, which differ in substrate chain length specificity and in susceptibility to inhibition by the antibiotic.

Acetates↗

Molecular properties of membrane lipids and activity of a membrane adenosine triphosphatase from Acholeplasma laidlawii B.

The effects of membrane lipid fluidity and phase state on enzyme activity were evaluated by careful analysis of Arrhenius plots of the Na+, Mg2+-dependent adenosine triphosphatase (referred to hereafter as ATPase) from Acholeplasma laidlawii B membranes and differential thermal analysis of the membrane lipid phase transitions. Arrhenius plots of the ATPase activity in membranes containing only liquid-crystalline lipids are gently curved and have the same form regardless of the membrane fatty liver composition. The absolute ATPase activity at temperatures well above the lipid phase transition temperature varies in a complex fashion with fatty acid composition. As the membrane enters its lipid phase transition, ATPase activity begins to fall off more rapidly than it does above the transition. Quantitative analysis of the results suggests that ATPase is reversibly inactivated when its vicinal lipids undergo a transition to a state of reduced plasticity at low temperatures. This transition is driven by the conversion of the bulk membrane lipid phase to the gel state, but it is less cooperative and occurs at lower temperatures than the bulk lipid transition. The results suggest that the familial "biphasic linear" Arrhenius plots reported for many membrane enzymes and transport systems may, in fact, have more complex shapes, analysis of which can furnish useful information about the behavior of the enzyme molecule or transport system in its membrane environment.

Acholeplasma laidlawii↗