Topological studies of the membrane component of the OleC ABC transporter involved in oleandomycin resistance in Streptomyces antibioticus.
The OleC ABC transporter of Streptomyces antibioticus is constituted by an ATP-binding protein (OleC) and a hydrophobic protein (OleC5). Here we present experimental evidence demonstrating that the OleC5 protein is an integral membrane protein and we propose a topological model for its integration into the membrane. This model is based on the generation of hybrid proteins between different regions of OleC5 and a Escherichia coli beta-lactamase (BlaM) and the determination of the minimal inhibitory concentrations to ampicillin in these constructions. Fusions were generated both by cloning specific fragments of oleC5 and by creating ExoIII nested deletions of the gene. In the topological model proposed there will be six alpha-helix transmembrane regions, two cytoplasmic and four periplasmic loops and a hydrophobic linker domain.
Olano
C
C
Departmento de Biologia Funcional, Universidad de Oviedo, Spain.
Rodriguez
A M
AM
Méndez
C
C
Salas
J A
JA
D018528:ATP-Binding Cassette Transporters / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D000595:Amino Acid Sequence; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001483:Base Sequence; D003001:Cloning, Molecular; D004274:DNA, Recombinant / Q000235:genetics; D004352:Drug Resistance, Microbial / Q000235:genetics; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009827:Oleandomycin / Q000494:pharmacology; D017433:Protein Structure, Secondary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013303:Streptomyces antibioticus / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D001618:beta-Lactamases / Q000235:genetics
D018528:ATP-Binding Cassette Transporters; D000900:Anti-Bacterial Agents; D004274:DNA, Recombinant; D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins; D001618:beta-Lactamases; D009827:Oleandomycin