Abstract
Ghosts of Micrococcus lysodeikticus contain a mannan that is not removed by intensive washing procedures. Purified mannan, isolated by extraction of whole cells with hot, aqueous phenol, binds to membranes in vitro. Mannan also binds to DEAE-cellulose and migrates toward the anode in neutral and sodium dodecyl sulfate disc gel electrophoresis. In aqueous solution mannan has an apparent molecular weight of 10-6, but in the presence of sodium dodecyl sulfate its apparent molecular weight is 50,000 to 100,000; removal of the detergent results in reaggregation. Purified mannan contains mannose, succinate, fatty acid, and glycerol in a ratio of 50:4.9:2.1:1.0. Treatment of mannan with mild base produces a neutral, hydrophilic polysaccharide of relatively low molecular weight that has no affinity for membranes. At least 90% of the reducing termini are blocked in a base-stable linkage. Based on these results a tentative structure is proposed for the mannan.
Highlights
When analyzed by disc gel electrophoresis in the presence of sodium dodecyl sulfate (Fig. 3B), intact mannan migrated toward the anode but, as in the neutral gel system, basehydrolyzed mannan was detected only at the top of the gel. These results suggested that intact mannan is acidic and binds sodium dodecyl sulfate and that both of these properties are lost after mild hydrolysis
Lipoteichoic acids have recently been isolated from a number of gram-positive bacteria [28,29,30]
Synthesis of the cell wall teiehoic xcid of StapkyloCOCCUSaureus II, polyribitol phosphate, is dependent on an acceptor which contains fatty acids as well as glycerol, phosphate, and glucose. This acceptor is indistinguishable in its composition and properties from the membrane-bound lipoteichoic acid of the same organism [31]
Summary
Solvent systems employed were: A, ethyl acetate-pyridine-water (12:5:4); B, ethyl acetate-pyridinewater (8:2: 1) ; C, isopropyl alcohol-concent,ratcd. NH&II (2: 1) ; D, benzene-methanol-formic acid (60:40:2); and E, isobutyric acid-concentrated NHdOH-water (57:4:39). Carbohydrates were detected by periodate-benzidine [19], and hydroxamic acids were visualized by spraying with 5yc FeC13 in ethanol [20]. Spcclroscopy-Optical density measrlrements were performed on a Gilford Spectrophotometer. W.iCochran of the Division of Bioohvsics at The Johns IToukins Universitv. 100 modified with a Digilab Pulse Unit and a Data General Computer to operate in the Fourier Transform mode
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