Sugar recognition mutants of the melibiose carrier of Escherichia coli: possible structural information concerning the arrangement of membrane-bound helices and sugar/cation recognition site.
Melibiose carrier mutants, isolated by growing cells on melibiose plus the non-metabolizable competitive inhibitor thiomethyl-beta-galactoside (TMG), were studied to determine sugar and cation recognition abnormalities. Most of the mutants show good transport of melibiose but have lost the recognition of TMG. In addition, most mutants show little or no transport of lactose. Cation recognition is also affected as all of these mutants have lost the ability to transport protons with melibiose. The amino acids causing these mutations were determined by sequencing the melB gene on the plasmid. The mutations were located on helices I, IV, VII, X and XI. We propose that these five helices are in proximity with each other and that they line the sugar/cation transport channel.
Ding
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PZ
Department of Cell Biology, Harvard Medical School, Boston, MA 02115, USA.
Botfield
M C
MC
Wilson
T H
TH
D001665:Binding Sites; D001667:Binding, Competitive; D001692:Biological Transport; D050260:Carbohydrate Metabolism; D002412:Cations; D002462:Cell Membrane / Q000201:enzymology; D004351:Drug Resistance; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D008553:Melibiose / Q000378:metabolism; D049990:Membrane Transport Modulators; D026901:Membrane Transport Proteins / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D008756:Methylgalactosides / Q000494:pharmacology; D009154:Mutation; D011487:Protein Conformation; D017494:Proton Pumps; D012964:Sodium / Q000378:metabolism; D027981:Symporters; D013862:Thiogalactosides / Q000494:pharmacology
D002412:Cations; D049990:Membrane Transport Modulators; D026901:Membrane Transport Proteins; D008756:Methylgalactosides; D017494:Proton Pumps; D027981:Symporters; D013862:Thiogalactosides; C009468:thiomethylgalactoside; C021548:melibiose permease; D008553:Melibiose; D012964:Sodium