Porin of Rhodobacter capsulatus: Biochemical and Functional Characterization

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Abstract
The major outer membrane protein of Rhodobacter capsulatus 37 b4 (capsule-free) was isolated. Strong porin-activity was observed after reconstitution into artificial lipid bilayer membranes with a single channel conductance of 3.15 nS in 1 m KCl. The porin migrated as a broad, single band (Mr above 90,000) on sodium dodecyl sulfate polyacrylamide gel electrophoresis and dissociated into a single species of polypeptides (Mr 36,000) on treatment with EDTA (10 mm at 30.degree. C, 20 min) or by heating (100.degree. C, 5 min). Analytical ultracentrifugation studies demonstrated the native porin to be a trimer. The monomers chromatofocused as a single, sharp peak on fast performance liquid chromatography and only one band, corresponding to an isoelectric point of about 4.0, was obtained on isoelectric focusing. Gas-phase sequencing of the 23 N-terminal residues revealed Glu-Val-Lys-Leu-Ser-Gly-Asp-Ala-Arg-Met-Gly-Val-Met-Tyr-Asn-Gly-Asp-Asp-X-Asn-Phe-Ser-Ser.