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Analysis of the topology of a membrane protein by using a minimum number of alkaline phosphatase fusions.

Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts 02115.

ABSTRACT

An approach to analyzing the topology of membrane proteins with alkaline phosphatase fusions is described. Precise fusions were constructed by using polymerase chain reaction at the C terminus of each hydrophilic region of the membrane protein. The disruption of topogenic signals is thereby minimized, and predictable anomalous results are avoided. The Escherichia coli MalG protein has been analyzed.

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