Highly efficient over-production in E. coli of YvcC, a multidrug-like ATP-binding cassette transporter from Bacillus subtilis.
ATP-binding cassette (ABC) transporters have often been refractory to over-expression. Using the C41(DE3) E. coli as a host strain, membrane vesicles highly enriched (>50%) in YvcC, a previously uncharacterized ABC transporter from Bacillus subtilis homologous to P-glycoprotein multidrug transporters, were obtained. The functionality of YvcC was assessed by its high vanadate-sensitive ATPase activity and its ability to transport a fluorescent drug, the Hoechst 33342.
Steinfels
Emmanuelle
E
Institut de Biologie et Chimie des Protéines, UMR 5086 CNRS/Université Claude Bernard Lyon I, 7 passage du Vercors, 69367, Lyon, France. jm.jault@ibcp.fr
Orelle
Cédric
C
Dalmas
Olivier
O
Penin
François
F
Miroux
Bruno
B
Di Pietro
Attilio
A
Jault
Jean-Michel
JM
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000096:biosynthesis; D018528:ATP-Binding Cassette Transporters / Q000096:biosynthesis*; D001412:Bacillus subtilis / Q000235:genetics*; D001425:Bacterial Outer Membrane Proteins / Q000096:biosynthesis*; D001562:Benzimidazoles / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000378:metabolism; D014638:Vanadates
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D018528:ATP-Binding Cassette Transporters; D001425:Bacterial Outer Membrane Proteins; D001562:Benzimidazoles; D005456:Fluorescent Dyes; D014638:Vanadates; C017807:bisbenzimide ethoxide trihydrochloride