Bacillus stearothermophilus lctB gene gives rise to functional K+ channels in Escherichia coli and in Xenopus oocytes.
We have cloned a small K+ channel subunit (LctB) of the gram-positive bacterium Bacillus stearothermophilus (B. stearo.). The B. stearo. LctB protein is only 134 amino acids long. The sequence contains a typical K+ channel P-domain with a K+ channel GYGD signature sequence and two hydrophobic, possibly membrane-spanning segments M1 and M2. Unexpectedly, LctB K+ channels exhibited properties which differed markedly from the ones reported for KcsA channels of the gram-positive bacterium Streptomyces lividans. LctB channels, when expressed in E. coli, were targeted to the outer membrane and not like KcsA channels to the inner membrane. After reconstitution in black lipid membrane, LctB channels mediated K+ currents at neutral pH. They were apparently not gated by pH like KcsA channels. Also, LctB cRNA produced functional LctB channels in the Xenopus oocyte expression system in marked contrast to KcsA. The results demonstrated that heterologous expression produced functional LctB channels both in E. coli and in Xenopus oocytes. It is proposed that bacterial LctB subunits can be properly handled by the Xenopus oocyte leading to the occurrence of functional LctB K+ channels in the oocyte plasma membrane.
Wolters
M
M
Zentrum für Molekulare Neurobiologie Hamburg, Institut für Neurale Signalverarbeitung, Germany.
Madeja
M
M
Farrell
A M
AM
Pongs
O
O
D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins; D001464:Barium / Q000494:pharmacology; D002586:Cesium / Q000494:pharmacology; D004553:Electric Conductivity; D004926:Escherichia coli / Q000378:metabolism; D005260:Female; D005798:Genes, Bacterial; D001411:Geobacillus stearothermophilus / Q000235:genetics*; D008081:Liposomes; D008969:Molecular Sequence Data; D009865:Oocytes / Q000378:metabolism; D018408:Patch-Clamp Techniques; D026902:Potassium Channel Blockers; D015221:Potassium Channels / Q000235:genetics / Q000378:metabolism*; D020836:Protein Structure, Quaternary; D011994:Recombinant Proteins / Q000378:metabolism; D014981:Xenopus
D001426:Bacterial Proteins; D008081:Liposomes; D026902:Potassium Channel Blockers; D015221:Potassium Channels; D011994:Recombinant Proteins; C107790:prokaryotic potassium channel; D002586:Cesium; D001464:Barium