Cysteines in the Shaker K+ channel are not essential for channel activity or zinc modulation.
We investigated whether the cysteine residues in Shaker potassium (K+) channels are essential for activation and inactivation gating or for modulation of activation gating by external zinc (Zn2+). Mutants of the Shaker K+ channel were prepared in which all seven cysteine residues were replaced (C-less). These changes were made in both wild-type Shaker H4 channels and in a deletion mutant (delta 6-46) lacking N-type ("fast") inactivation. Replacement of all cysteines left most functional properties of the K+ currents unaltered. The most noticeable difference between the C-less and wild-type currents was the faster C-type inactivation of the C-less channel which could be attributed largely to the mutation of Cys462. This is consistent with the effects of previously reported mutations of nearby residues in the S6 region. There were also small changes in the activation gating of C-less currents. Modulation by external Zn2+ of the voltage dependence and rate of activation gating is preserved in the C-less channels, indicating that none of the cysteines in the Shaker K+ channel plays an important role in Zn2+ modulation.
Department of Neurobiology, Massachusetts General Hospital, Boston 02114.
L M
LM
Boland
M E
ME
Jurman
G
G
Yellen
D000818:Animals; D055592:Biophysical Phenomena; D001703:Biophysics; D002460:Cell Line; D003545:Cysteine / Q000737:chemistry / Q000235:genetics; D006801:Humans; D015640:Ion Channel Gating / Q000187:drug effects; D007700:Kinetics; D016297:Mutagenesis, Site-Directed; D009865:Oocytes / Q000378:metabolism; D010455:Peptides / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D026902:Potassium Channel Blockers; D015221:Potassium Channels / Q000235:genetics / Q000378:metabolism*; D017384:Sequence Deletion; D051637:Shaker Superfamily of Potassium Channels; D014982:Xenopus laevis; D015032:Zinc / Q000494:pharmacology
D010455:Peptides; D026902:Potassium Channel Blockers; D015221:Potassium Channels; C076413:Shaker B potassium channel polypeptide; D051637:Shaker Superfamily of Potassium Channels; D015032:Zinc; D003545:Cysteine