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The membrane topology of the Rhizobium meliloti C4-dicarboxylate permease (DctA) as derived from protein fusions with Escherichia coli K12 alkaline phosphatase (PhoA) and β-galactosidase (LacZ)

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Abstract

The Rhizobium meliloti dctA gene encodes the C4-dicarboxylate permease which mediates uptake of C4-dicarboxylates, both in free-living and symbiotic cells. Based on the hydrophobicity of the DctA protein, 12 putative membrane spanning regions were predicted. The membrane topology was further analysed by isolating in vivo fusions of DctA to Escherichia coli alkaline phosphatase (PhoA) and E. coli β-galactosidase (LacZ). Of 10 different fusions 7 indicated a periplasmic and 3 a cytoplasmic location of the corresponding region of the DctA protein. From these data a two-dimensional model of DctA was constructed which comprised twelve transmembrane α-helices with the amino-terminus and the carboxy-terminus located in the cytoplasm. In addition, four conserved amino acid motifs present in many eukaryotic and prokaryotic transport proteins were observed.

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Authors and Affiliations

  1. Lehrstuhl fur Genetik, Fakultät für Biologie, Universität Bielefeld, D-33501, Bielefeld, Germany

    Doris Jording & Alfred Pühler

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  1. Doris Jording
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  2. Alfred Pühler
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Communicated by J. Lengeler

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Jording, D., Pühler, A. The membrane topology of the Rhizobium meliloti C4-dicarboxylate permease (DctA) as derived from protein fusions with Escherichia coli K12 alkaline phosphatase (PhoA) and β-galactosidase (LacZ). Molec. Gen. Genet. 241, 106–114 (1993). https://doi.org/10.1007/BF00280207

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  • DOI: https://doi.org/10.1007/BF00280207

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