Hancock W S, Harding D R, Barling P M, Sparrow J T
Int J Pept Protein Res. 1985 Jul;26(1):105-8. doi: 10.1111/j.1399-3011.1985.tb03184.x.
Apolipoproteins C-I and A-I were radioactively labelled with tritium by (3H)-methyl exchange. The methionine residues were first methylated with (3H)-methyl iodide at pH 4 and the reaction products were purified by gel filtration and cation exchange chromatography. The products were then demethylated with 2-mercaptoethanol (6 M) at pH 8.6 to regenerate the apolipoproteins in an unmodified but tritiated form. The specific radioactivity for apolipoprotein C-I and A-I was 3.5 X 10(6) and 1.5 X 10(7) dpm/pmol respectively. The properties of (3H)-apolipoprotein C-I were examined by reversed phase HPLC and by incorporation into very low density lipoproteins (VLDL).
载脂蛋白C-I和A-I通过(3H)-甲基交换用氚进行放射性标记。首先在pH 4下用(3H)-甲基碘将甲硫氨酸残基甲基化,反应产物通过凝胶过滤和阳离子交换色谱法纯化。然后在pH 8.6下用2-巯基乙醇(6M)将产物去甲基化,以未修饰但含氚的形式再生载脂蛋白。载脂蛋白C-I和A-I的比放射性分别为3.5×10(6)和1.5×10(7)dpm/pmol。通过反相高效液相色谱法并将其掺入极低密度脂蛋白(VLDL)中来检测(3H)-载脂蛋白C-I的性质。