1. Department of Anesthesiology, Washington University School of Medicine, Box 8054, St. Louis, Missouri 63110, USA

Correspondence to: Christopher J. Lingle Correspondence and requests for materials should be addressed to C.J.L. (e-mail: Email: clingle@morpheus.wustl.edu).

Abstract

Large conductance, Ca²⁺- and voltage-activated K⁺ channels (BK) respond to two distinct physiological signals—membrane voltage and cytosolic Ca²⁺ (refs 1, 2). Channel opening is regulated by changes in Ca²⁺ concentration spanning 0.5 M to 50 mM (refs 2–5), a range of Ca²⁺ sensitivity unusual among Ca²⁺-regulated proteins. Although voltage regulation arises from mechanisms shared with other voltage-gated channels^{6, 7, 8}, the mechanisms of Ca²⁺ regulation remain largely unknown. One potential Ca²⁺-regulatory site, termed the 'Ca²⁺ bowl', has been located to the large cytosolic carboxy terminus^{9, 10, 11}. Here we show that a second region of the C terminus, the RCK domain (regulator of conductance for K⁺ (ref. 12)), contains residues that define two additional regulatory effects of divalent cations. One site, together with the Ca²⁺ bowl, accounts for all physiological regulation of BK channels by Ca²⁺; the other site contributes to effects of millimolar divalent cations that may mediate physiological regulation by cytosolic Mg²⁺ (refs 5, 13). Independent regulation by multiple sites explains the large concentration range over which BK channels are regulated by Ca²⁺. This allows BK channels to serve a variety of physiological roles contingent on the Ca²⁺ concentration to which the channels are exposed^{14, 15}.