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pubmed:abstractText |
A protein conferring passive chloride permeability was isolated from a N-octylglucoside solubilized extract of partially purified H(+)-transporting osteoclast cell membranes. Purification was achieved by binding of solubilized protein to an amine-linked 4,4'-diisothiocyanatostilbene-2,2'-disulfonate (DIDS) Sepharose 4B column and elution with 50 mM KCl. A major protein, with MR = 60 kD on 10% SDS-PAGE, was obtained, which was further purified to homogeneity by HPLC gel filtration. This protein introduced 36Cl- permeability when reconstituted in phospholipid membranes by equilibrium dialysis. The Cl- transport recovered in reconstituted membranes retained sensitivity to DIDS confirming the identity of the isolated protein as a stilbene-sensitive chloride channel.
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