Abstract
The separation of outer and cytoplasmic membrane fractions has been difficult to achieve with pseudomonads. A new procedure is described in which Pseudomonas facilis is treated with methyl picolinimidate, subsequently with buffered sucrose, and finally with lysozyme-EDTA to yield spheroplasts; spheroplast-derived envelopes can then be resolved by density gradient centrifugation into several components only one of which is more dense in the presence of Ni 2+. This latter component, corresponding to the cell wall fraction, contains about 2% of the succinate dehydrogenase, a cytoplasmic membrane marker. Analogous experiments with erythrocytes establish that spectrin, a protein known to be located on the inner membrane surface, is not labeled by the methyl picolinimidate.
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