A secretion vector, pVN1, expressing the [NiFe] hydrogenase signal peptide of Hildenborough fused to β-lactamase from was constructed in order to study the unusual characteristics of hydrogenase signal peptides, which share a strictly conserved sequence, the consensus box: R-R-X-F-X-K. Although the hydrogenase signal peptide-β-lactamase fusion protein was processed much more slowly than the fusion of β-lactamase with its own signal peptide, the system mimicked several features expected for hydrogenase biosynthesis in , including increased export under anaerobic conditions. Site-directed mutagenesis of R(-28), the first arginine residue of the consensus box, to a glutamate completely inhibited export and processing of the fusion protein. The same mutation of R(-33), located outside the consensus box, had almost no effect. The data indicate a specific role for the consensus box sequence in the export mechanism for hydrogenase.


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