Effects of protein modification procedures on the interaction between 25-hydroxyvitamin D and the human plasma binding protein for vitamin D and its metabolites.
Effects of protein modification procedures on the interaction between 25-hydroxyvitamin D and the human plasma binding protein for vitamin D and its metabolites.
复制标题
蛋白质修饰程序对 25-羟基维生素 D 与维生素 D 及其代谢物的人血浆结合蛋白之间相互作用的影响。
DOI：
10.1021/bi00523a035
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发表时间：
1981
期刊：
影响因子：
2.9
通讯作者：
Goodman,DS
中科院分区：
文献类型：
--
作者：
Kawakami,M;Goodman,DS
材料与方法DBP的分离。通过与本实验室先前描述的方法类似的方法从合并的人血浆中分离DBP（Imawari等人，1976年）。针对单特异性抗人DBP抗血清（Imawari & Goodman，1977）的免疫反应性在DBP分离过程中用作DBP的标志物。本文报道的用于分离DBP的程序顺序包括在SP-Sephadex上柱层析，然后在DEAE-纤维素上柱层析，然后在固定的Cibacron Blue F3 GA（Pierce）上柱层析，然后在Sephacryl S-200上凝胶过滤。在SP-Sephadex上层析期间，全DBP [即，含有1的DBP]使用的缩写：DBP，维生素D及其代谢物的结合蛋白; Gc蛋白，组特异性组分蛋白; 25（OH）D，25-羟基维生素D; 1，25（OH）2D，1，25-二羟基维生素D; 24，25-（OH）2D，24，25-二羟基维生素D; DTE，二硫代戊糖醇; DTT，二硫苏糖醇; PBS，磷酸盐缓冲液; DEAE，二乙基氨基乙基; Tris，三（羟甲基）氨基甲烷; EDTA，乙二胺四乙酸。
Materials and MethodsIsolation of DBP. DBP was isolated from pooled human plasma by methods similar to those described previouslyfrom this laboratory (Imawari et al., 1976). Immunoreactivity against monospecific anti-human DBP antiserum (Imawari & Goodman, 1977) was used as a marker for DBP during the course of its isolation. The sequence of procedures used to isolate DBP for the studies reported here included column chromatography on SP-Sephadex, then on DEAE-cellulose, and then on immobilized Cibacron Blue F3GA (Pierce), followed by gel filtration on Sephacryl S-200. During chromatography on SP-Sephadex, holo-DBP [ie, DBP containing1 Abbreviations used: DBP, binding protein for vitamin D and its metabolites; Gc protein, group-specific component protein; 25 (OH) D, 25-hydroxyvitamin D; l, 25 (OH) 2D, 1, 25-dihydroxyvitamin D; 24, 25-(OH) 2D, 24, 25-dihydroxyvitamin D; DTE, dithioerythritol; DTT, dithiothreitol; PBS, phosphate-buffered saline; DEAE, diethylaminoethyl; Tris, tris (hydroxymethyl) aminomethane; EDTA, ethylenediaminetetraacetic acid.