19F and 31P NMR spectroscopy of G protein alpha subunits. Mechanism of activation by Al3+ and F-.
作者： T Higashijima , M P Graziano , H Suga , M Kainosho , A G Gilman
DOI: 10.1016/S0021-9258(19)67806-3
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摘要: 19F and 31P NMR spectroscopy was used to study the mechanism of activation alpha subunits guanine nucleotide-binding regulatory proteins (G proteins) by Al3+, Mg2+, F-. spectra solutions containing F- showed a characteristic peak at -10 ppm. Addition GDP-bound form either two G protein alpha) resulted in appearance an additional -29 or -30 This not observed with guanosine 5'-3-O-(thio)triphosphate-G GDP alone. Titration indicated that each molecule binds 3-5 molecules (Kd = 0.47 mM), single Al3+ much less than 0.1 Mg2+ ion about mM). Replacement Mn2+ caused dramatic broadening signal, indicating metal proximity protein-bound (less 1 nm). GDP-G peaks -2 -8.6 ppm, corresponding beta- alpha-phosphoryl groups GDP, respectively. Binding upfield shift 6 ppm for beta-phosphoryl signal no change signal. These observations indicate Mg2+.GDP.AlF3-5 mimics Mg2+.GTP its capacity activate subunits.
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