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Inositol Polyphosphate Receptor and Clathrin Assembly Protein AP-2 are Related Proteins that Form Potassium-Selective Ion Channels in Planar Lipid Bilayers
Ist Teil von
Proceedings of the National Academy of Sciences - PNAS, 1992-10, Vol.89 (19), p.8976-8980
Ort / Verlag
Washington, DC: National Academy of Sciences of the United States of America
Erscheinungsjahr
1992
Quelle
Free E-Journal (出版社公開部分のみ)
Beschreibungen/Notizen
We have previously described an inositol polyphosphate receptor (IPxRec), purified from detergent-solubilized bovine cerebellum microsomes, that displays potassium ion channel activity in planar lipid bilayers. We now find that the IPxRec is closely related to clathrin assembly protein AP-2. The IPxRec and AP-2 purified from bovine brain clathrin-coated vesicles share several structural and functional features: (i) similar subunit composition; each has four major polypeptides that have similar mobility (Mrvalues of 111,000, 100,000, 50,000, and 17,000) and relative intensity by SDS/PAGE analysis; (ii) similar size as studied by molecular sieve chromatography (Mr400,000); (iii) identical N-terminal amino acid sequences for the Mr50,000 subunits and Mr111,000/100,000 doublets; (iv) immunoreactivity of the AP-2 Mr111,000/100,000 doublet to polyclonal antibodies affinity purified against the doublet proteins of the IPxRec; (v) display of the in vitro diagnostic feature of assembly proteins-i.e., they induce the assembly of clathrin cages; and (vi) ion channel activity selective for potassium ions with the same unitary conductance when incorporated into planar lipid bilayers. One difference was found. AP-2 channels were not blocked by inositol 1,3,4,5-tetraphosphate as reported for IPxreceptor channels. These studies suggest a possible connection between the IPxsignaling pathways and receptor-mediated endocytosis.