Different foci for the regulation of the activity of the KefB and KefC glutathione-gated K+ efflux systems.
KefB and KefC are glutathione-gated K+ efflux systems in Escherichia coli, and the proteins exhibit strong similarity at the level of both primary sequence and domain organization. The proteins are maintained closed by glutathione and are activated by binding of adducts formed between glutathione and electrophiles. By construction of equivalent mutations in each protein, this study has analyzed the control over inactive state of the proteins. A UV-induced mutation in KefB, L75S, causes rapid spontaneous K+ efflux but has only a minor effect on K+ efflux via KefC. Similarly amino acid substitutions that cause increased spontaneous activity in KefC have only small effects in KefB. Exchange of an eight amino acid region from KefC (HALESDIE) with the equivalent sequence from KefB (HELETAID) has identified a role for a group of acidic residues in controlling KefC activity. The mutations HELETAID and L74S in KefC act synergistically, and the activity of the resultant protein resembles that of KefB. We conclude that, despite the high degree of sequence similarity, KefB and KefC exhibit different sensitivities to the same site-specific mutations.
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Department of Molecular and Cell Biology, University of Aberdeen, Institute of Medical Sciences, Foresterhill, Aberdeen AB25 2ZD, United Kingdom.
Booth
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D000595:Amino Acid Sequence; D017920:Antiporters / Q000235:genetics / Q000502:physiology* / Q000528:radiation effects; D001426:Bacterial Proteins / Q000235:genetics / Q000502:physiology* / Q000528:radiation effects; D003001:Cloning, Molecular; D017124:Conserved Sequence; D004926:Escherichia coli / Q000235:genetics; D029968:Escherichia coli Proteins; D005978:Glutathione / Q000502:physiology*; D015640:Ion Channel Gating / Q000502:physiology*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D015221:Potassium Channels / Q000235:genetics / Q000502:physiology* / Q000528:radiation effects; D027961:Potassium-Hydrogen Antiporters; D011765:Pyruvaldehyde / Q000494:pharmacology; D016415:Sequence Alignment; D013329:Structure-Activity Relationship; D014466:Ultraviolet Rays
D017920:Antiporters; D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D015221:Potassium Channels; D027961:Potassium-Hydrogen Antiporters; C069287:KefC protein, E coli; D011765:Pyruvaldehyde; D005978:Glutathione