The Azotobacter vinelandii gene algJ encodes an outer-membrane protein presumably involved in export of alginate
The algJ gene from Azotobacter vinelandii was cloned using a labelled RNA probe representing the coding region of the algE gene from Pseudomonas aeruginosa. DNA sequencing revealed an ORF of 1452 bp encoding a protein of 484 amino acid residues with a calculated molecular mass of 54611 Da. An RNA probe corresponding to algE was also used for Southern hybridization of chromosomal DNA, which showed that algE-related DNA sequences are also present in the alginate-producing phytopathogen species Pseudomonas marginalis and Pseudomonas syringae pv. glycinea. The coding region of algJ was subcloned in the expression vector pT7-7, leading to a corresponding gene product with an apparent molecular mass of 54 kDa which could be identified in the outer membrane (OM) of Escherichia coli BL21(DE3). Additionally, a cross-reacting protein with the same molecular mass was also found in the OM of A. vinelandii using an anti-AlgE antiserum. The derived amino acid sequence of AlgJ shared approximately 52% identity with AlgE from P. aeruginosa. The hydrophilicity profile as well as the amphipathicity of regions in the amino acid sequence of AlgJ showed significant similarities to AlgE. Based on these data, a topological model of AlgJ was created with the aid of known structures of outer-membrane proteins. This model presents AlgJ as a ß-barrel containing 18 ß-strands inserted in the OM.
Biochemistry and Cell Biology not elsewhere classified