Gw. Abbott et al., CONFORMATIONAL-CHANGES IN A MAMMALIAN VOLTAGE-DEPENDENT POTASSIUM CHANNEL INACTIVATION PEPTIDE, Biochemistry, 37(6), 1998, pp. 1640-1645
Fast inactivation is restored in inactivation deletion mutant voltage-
gated potassium (K-v) channels by application of synthetic inactivatio
n 'ball' peptide. Using Fourier transform infrared and circular dichro
ism spectroscopy, we have investigated the structure of synthetic K(v)
3.4 channel ball peptide, in a range of environments relevant to the f
unction of the ball domain. The ball peptide contains no alpha-helix o
r beta-sheet in reducing conditions in aqueous solution, but when coso
lubilized with anionic lipid or detergent in order to mimic the enviro
nment which the ball domain encounters during channel inactivation, th
e ball peptide adopts a partial beta-sheet structure. Oxidation of the
K(v)3.4 ball peptide facilitates formation of a disulfide bond betwee
n Cys(6) and Cys(24) and adoption of a partial beta-sheet structure in
aqueous solution; the tendency of the oxidized ball peptide to adopt
beta-sheet is generally greater than that of the reduced ball peptide
in a given environment. THREADER modeling of the K(v)3.4 ball peptide
structure predicts a beta-hairpin-like conformation which corresponds
well to the structure suggested by spectroscopic analysis of the ball
peptide in its cyclic arrangement, A V7E mutant K(v)3.4 ball peptide a
nalogue of the noninactivating Shaker B L7E mutant ball peptide cannot
adopt beta-structure whatever the environment, and regardless of oxid
ation state. The results suggest that the K(v)3.4 ball domain undergoe
s a conformational change during channel inactivation and may implicat
e a novel regulatory role for intramolecular disulfide bond formation
in the K(v)3.4 ball domain in vivo.