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J. Biol. Chem., Vol. 279, Issue 21, 22461-22468, May 21, 2004

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A Novel Human Cl^- Channel Family Related to Drosophila flightless Locus^*

Makoto Suzuki and Atsuko Mizuno

From the Department of Pharmacology, Jichi Medical School 3311-1, Yakushiji, Minamikawachi, Tochigi 329-0498, Japan

Large conductance chloride (maxi-Cl^-) currents have been recorded in some cells, but there is still little information on the molecular nature of the channel underlying this conductance. We report here that tweety, a gene located in Drosophila flightless, has a structure similar to those of known channels and that human homologues of tweety (hTTYH1–3) are novel maxi-Cl^- channels. hTTYH3 mRNA was found to be distributed in excitable tissues. The whole cell current of hTTYH3 was large enough to be discriminated from the control but emerged only after treatment with ionomycin. Analysis of pore mutants suggested that positively charged amino acids contributed to anion selectivity. Like a maxi-Cl^- channel in situ, the hTTYH3 single channel showed 26-picosiemen linear current voltage, complex kinetics, 4,4'-diisothiocyanato-stilbene-2,2'-disulfonic acid sensitivity, subconductance, and the permeability order of I^- > Br^- > Cl^-. Similarly, hTTYH2 encoded an ionomycin-induced maxi-Cl^- channel, but TTYH1 encoded a Ca²⁺-independent and swelling-activated maxi-Cl^- channel. Therefore, the hTTYH family encoded maxi-Cl^- channels of mammals. Further studies on the hTTYH family should lead to the elucidation of physiological and pathophysiological roles of novel Cl^- channel molecules.

Received for publication, December 17, 2003 , and in revised form, March 8, 2004.

The nucleotide sequence(s) reported in this paper has been submitted to the DDBJ/GenBank^TM/EBI Data Bank with accession number(s) AB162931, BC005168, AB162929, AB162930, and NM020659.

^* The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

To whom correspondence should be addressed: Dept. of Pharmacology, Jichi Medical School 3311-1, Yakushiji, Minamikawachi, Tochigi 329-0498, Japan. Tel.: 81-285-58-7326; Fax: 81-285-44-5541; E-mail: macsuz{at}jichi.ac.jp.

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