The Major Proteins of the <em>Escherichia coli</em> Outer Cell Envelope Membrane.

A procedure is described that from one batch of cells allows the isolation of all major proteins of the outer cell envelope membrane of Escherichia coli B/r. The method involves differential extraction of cell envelopes with ionic and non-ionic detergents with and without Mg²⁺ present, an the proteins are finally separated by molecular sieve chromatography in the presence of sodium dodecylsulfate. From 200 g cell paste in ten days (including the five days chromatography) ≈ 120 mg protein I (molecular weight ≈ 38000), ≈ 110 mg protein II* (molecular weight ≈ 33000), ≈ 50 mg protein III (molecular weight ≈ 17000), and ≈ 30 mg protein IV (molecular weight ≈ 7000) are obtained in pure state, and these yields are near the expected ones assuming quantitative recoveries. Protein II* is a heat-modifiable protein (perhaps due to complete unfolding and/or binding of sodium dodecylsulfate only at higher temperatures), and the isolated protein is completely in its unmodified form. Protein IV, Braun's lipoprotein, in the cell envelope exists in two forms, one covalently bound to the murein layer and the other not. The isolated protein IV represents the free form of the protein that so far had not been isolated; its protein part does not differ substantially from that of the bound form. [ABSTRACT FROM AUTHOR]