Abstract:
:A model for the topology of the PhoE porin has been proposed according to which the polypeptide traverses the outer membrane sixteen times mostly as amphipathic beta-sheets, thereby exposing eight loops at the cell surface. Until now, no evidence has been obtained for the surface exposure of the third loop. Recently, the structure of porin of Rhodobacter capsulatus has been determined. The proposed model of PhoE is very similar to the structure of the R. capsulatus porin, which has an 'eyelet' region, extending into the interior of the pore. The proposed third external loop of PhoE might form a similar 'eyelet' region. To determine the location of the predicted third external loop of PhoE, multiple copies of an oligonucleotide linker encoding an antigenic determinant of VP1 protein of foot-and-mouth disease virus (FMDV) were inserted. All hybrid proteins were properly inserted in the outer membrane. The monoclonal antibody MA11, directed against the linear FMDV epitope, was able to bind only to intact cells expressing a hybrid PhoE protein with at least three copies of the FMDV epitope present. Antibiotic sensitivity tests and single-channel conductance measurements revealed that the insertions influenced the channel size. These results are consistent with a location of the third loop of PhoE within the pore channel.
journal_name
Mol Microbioljournal_title
Molecular microbiologyauthors
Struyvé M,Visser J,Adriaanse H,Benz R,Tommassen Jdoi
10.1111/j.1365-2958.1993.tb01104.xsubject
Has Abstractpub_date
1993-01-01 00:00:00pages
131-40issue
1eissn
0950-382Xissn
1365-2958journal_volume
7pub_type
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