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- W2189341007 abstract "Withprocedures developed previously inthis laboratory, thevarious layers ofthe cell wallofa gram-negative bacterium, a marinepseudomonad (ATCC 19855), were removed completely giving rise totrueprotoplasts. Membranes were isolated fromtheprotoplasts formed. After treatment withribonuclease, deoxyribonuclease, andwashing, themembranes isolated were shownbyelectron microscopy and chemical analysis tobeessentially freefrombothwallmaterial andcytoplasmic constituents. Themembranes gave rise toa single compactbandina sucrosedensity gradient. Allofthelipid andprotein were found tobeassociated inthemembrane band.Analysis showed themembranes tocontain 30.5%lipid (78%ofwhich was phospholipid), 62.8%protein, and2% carbohydrate. Thepredominant phospholipid present was phosphatidylethanolamine with a lesser amount ofdiphosphatidylglycerol andtracesofunidentified compounds. Inmostprevious attempts toisolate purified andhomogenous cytoplasmic membranes from gram-negative bacteria, onlylimited success has beenachieved. Forgram-positive bacteria, stabilized protoplasts areformed withfacility andexhibit minimal, ifany,remaining cell wallcomponents(10,28,32).Cellmembranes arethen readily obtained andpurified fromtheprotoplasts ofgram-positive cells (28). Thecell wallcytoplasmic membrane complex (cell envelope) ofgram-negative bacteria presents amorecomplicated situation. Inaddition toan innercytoplasmic membrane, another outer membrane structure orcell walldouble track has beenobserved (16,21). A rigid peptidoglycan layer hasalsobeendemonstrated between the outerdouble tracklayer andtheplasma membrane(22). Although osmotically fragile spheroplasts canreadily beformedfrommostgramnegative bacteria (18), atthistimethere have beennoreports fromother laboratories ofthe successful removal ofallofthecell wallmaterial fromagram-negative bacterium andtheformation ofatrueprotoplast fromsuchanorganism. Previous studies inthis laboratory haveelucidated asystem forthecomplete removal ofallof thecell wallmaterial fromagram-negative marinepseudomonad (7,8).Fromthestable, true protoplasts formed bythetechnique developed, a procedure hasnowbeendevised fortheisolation andpurification ofhomogeneous cytoplasmic 1160 membranes fromtheorganism. Thepurity and homogeneity ofthese cytoplasmic membranes havebeenestablished andtheir chemical composition hasbeendetermined." @default.
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- W2189341007 title "Isolation andChemical Composition ofthe Cytoplasmic Membraneofa Gram-Negative Bacterium" @default.
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