Export of hybrid proteins FhuA'-'LacZ and FhuA'-'PhoA to the cell envelope of Escherichia coli K-12.
The fhuA gene of Escherichia coli K-12 encodes an outer membrane protein that acts as the ferrichrome-iron(III) receptor. To determine the export signals and sorting information within FhuA, gene fusions of fhuA'-'lacZ and fhuA'-'phoA were constructed. Although a FhuA'-'LacZ hybrid protein was detected in the Triton X-100-insoluble fraction of the cell envelope, direct immunoelectron microscopic observation showed that this protein remained in the cytoplasm. FhuA'-'PhoA hybrid proteins were all exported across the cytoplasmic membrane. Those hybrids containing up to 88 amino acids of FhuA (FhuA88) fused to PhoA were released along with other periplasmic proteins. Hybrids containing 180 or more amino acids of FhuA (FhuA180) fused to PhoA were associated with the outer membrane. It is proposed that some information inherent in the sequences between FhuA88 and FhuA180 confers stable association with the outer membrane.
Coulton
J W
JW
Department of Microbiology and Immunology, McGill University, Montreal, Quebec, Canada.
Reid
G K
GK
Campana
A
A
D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D002458:Cell Fractionation; D002462:Cell Membrane / Q000032:analysis / Q000378:metabolism; D003001:Cloning, Molecular; D003593:Cytoplasm / Q000032:analysis; D004926:Escherichia coli / Q000032:analysis / Q000235:genetics* / Q000378:metabolism / Q000648:ultrastructure; D005798:Genes, Bacterial; D007118:Immunoassay; D007150:Immunohistochemistry; D008854:Microscopy, Electron; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism*
D001425:Bacterial Outer Membrane Proteins; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins