TuCa1, an ascidian homolog of L-type Ca channel alpha(1)-subunit, has many critical sites required for binding 1,4-dihydropyridines (DHPs), but is insensitive to DHPs and methyl 2,5-dimethyl-4-[2-(phenylmethyl)benzoyl]-1H-pyrrole-3-carboxylate (FPL-64176). We have substituted Ser for Ala(1016) at the P-region of domain III in TuCa1 (TuCa1/A1016S) and functionally expressed the channel in Xenopus oocyte along with rabbit alpha(2)/delta and beta(2b). TuCa1/A1016S has gained DHP sensitivity as high as that of a mammalian neuronal L-type Ca channel (rbCII), but remained resistant to FPL-64176. These results reinforce the view that Ser(1016) in TuCa1/A1016S participates in DHP binding, but there exist other novel sites that fully acquire sensitivity to FPL-64176.
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